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Danlami, Mohammed Bashar, Basiru Aliyu, and Grace Samuel. "INCIDENCE OF RIFAMPICIN-RESISTANCE PRESUMPTIVE M. TUBERCULOSIS CASES AMONG OUTPATIENTS IN KEBBI STATE, NIGERIA." African Journal of Infectious Diseases 15, no. 1 (December 15, 2020): 47–52. http://dx.doi.org/10.21010/ajid.v15i1.6.
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Background: The present study determined the incidence of rifampicin resistance M. tuberculosis among outpatients at the General Hospital Yauri, Kebbi State, Nigeria. Materials and Methods: The study is a cross-sectional study conducted from February 2018 to October 2019. Sociodemographic data were collected from hospital registration books. Rifampicin resistance M. tuberculosis was detected using GeneXpert Model GX-IV following manufacturers' instruction. Descriptive statistics and logistic regression were computed using SPSS version 20. The results were presented as odds ratios with associated 95% confidence intervals, and P-value at 0.05. Result: Of the 837 samples, 65.8% (551/837) were males, and 34.2% (286/837) females, 11.4% (95/837) HIV-seropositive. M. tuberculosis was detected in 15.5% (130/837), of which 116/130 (89.23%) were males and 14/130 (10.77%) females. M. tuberculosis¬-HIV coinfection was detected in 9.47% (9/95) of HIV positive. Rifampicin resistance was observed in 1.3% (11/837), 7.7% (10/130) in M. tuberculosis patients and 1.05% (1/94) in HIV seropositive. In logistic regression, the odds ratio for having a rifampicin-resistant M. tuberculosis was 0.49 (0.15-1.54) for > 30 years; taking <30 years as the reference value, 1.02 (1.00-1.03) for male; taking female as the reference value, and 0.78 (0.09-6.15) for HIV positive, taking negative as the reference value. Conclusion: This study reported the current incidence rate of rifampicin-resistant M. tuberculosis at the General Hospital Yelwa Yauri, Kebbi State, Nigeria, among presumptive TB patients. Patients diagnosed with rifampicin-resistant M. tuberculosis were predominantly male adults. Thus, frequent screening is vital for surveillance and reduces the risk of transmission and spread of M. tuberculosis infections.
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de la Iglesia, Agustina I., Emma J. Stella, and Héctor R. Morbidoni. "Comparison of the Performances of Two In-House Rapid Methods for Antitubercular Drug Susceptibility Testing." Antimicrobial Agents and Chemotherapy 53, no. 2 (November 17, 2008): 808–10. http://dx.doi.org/10.1128/aac.00960-08.
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ABSTRACT Resistance to rifampin (rifampicin), isoniazid, and streptomycin of 69 Mycobacterium tuberculosis isolates was analyzed by an in-house method based on mycobacteriophage D29 and a colorimetric micromethod. Both methods showed sensitivity and specificity values ranging from 93% to 100%. These simple methods offer an option for drug resistance assessment of M. tuberculosis.
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АХМЕТОВА, А. Ж., Ж. М. АБИЛОВА, Г. Н. ЖАРЫЛҚАСЫН, А. Р. АКИЛЬЖАНОВА, and У. А. КОЖАМКУЛОВ. "ҚАЗАҚСТАНДА ТАРАЛҒАН РИФАМПИЦИН-ТӨЗІМДІ M. TUBERCULOSIS КЛИНИКАЛЫҚ ИЗОЛЯТТАРЫНЫҢ ГЕНЕТИКАЛЫҚ СИПАТТАМАСЫ." Vestnik, no. 1(64) (April 14, 2023): 53–65. http://dx.doi.org/10.53065/j5985-7576-8270-u.
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Лекарственно-устойчивый туберкулез остается актуальной проблемой в здравоохранении Казахстана. Казахстан одна из стран в мире с высоким количеством мультирезистентного туберкулеза, характеризующегося устойчивостью к основным антибиотикам первого ряда – рифампицину и изониазиду. Цель данной работы: определить биологическое разнообразие и охарактеризовать мутации в rpoB гене рифампицин-устойчивых клинических изолятов M. tuberculosis из Казахстана. В данной работе от больных с легочным туберкулезом было собрано 115 клинических изолятов M. tuberculosis, устойчивых к противотуберкулезному препарату первого ряда - рифампицину. Генотипирование всех 115 рифампицин-резистентных изолятов было проведено методом MIRU-VNTRс использованием 15 локусов. ДНК-секвенирование RRDR региона rpoB гена всех 115 рифампицин-устойчивых образцов M. tuberculosis было выполнено на секвенаторе ABI 3730 DNA Analyzer (Applied Biosystems). Согласно результатам MIRU-VNTR генотипирования, 86,1% образцов среди рифампицин-устойчивых изолятов были отнесены к семейству Beijing M. tuberculosis. Анализ нуклеотидных последовательностей всех устойчивых к рифампицину изолятов показал преобладание мутации в 531 кодоне rpoB гена с аминокислотной заменой серина на лейцин Ser531Leu в 82,6% случаев. В данной работе семейство Beijing M. tuberculosis было ассоциировано с мутацией Ser531Leu в 531 кодоне гена (p<0.0001), а генотип LAM M. tuberculosis был связан с мутацией His526Leu в 526 кодоне гена (p<0.0001). Дәріге-төзімді туберкулез Қазақстанның денсаулық сақтау саласында өзекті мәселе болып қалуда. Қазақстан бірінші қатардағы негізгі антибиотиктерге – рифампицин мен изониазидке төзімділікпен сипатталатын мультирезистентті туберкулез көрсеткіші жоғары мемлекеттердің бірі. Зерттеу жұмысының мақсаты: Қазақстанда таралған рифампицинге-төзімді M. tuberculosis клиникалық изоляттарының rpoB генінде мутацияларды сипаттау және олардың биотүрлігін анықтау. Аталған зерттеу жұмысында өкпе туберкулезі науқастарынан туберкулезге қарсы бірінші қатардағы препарат – рифампицинге төзімді 115 M. tuberculosis клиникалық изоляттары жиналды. Барлық 115 рифампицинге резистентті изоляттар MIRU-VNTR әдісімен 15 локус бойынша генотиптелді. 115 рифампицинге-төзімді M. tuberculosisүлгілерінің rpoB генінің RRDR аймағын ДНҚ-секвенирлеу ABI 3730 DNA Analyzer (Applied Biosystems) секвенаторында өткізілді. MIRU-VNTR генотиптеу нәтижелеріне сәйкес, рифампицин-төзімді изоляттардың арасында 86,1% жағдайында үлгілер Beijing M. tuberculosisтұқымдасына жатты. Рифампицинге төзімділігі бар барлық изоляттардың нуклеотидтік тізбектерінің анализі 82,6% жағдайында rpoB генінің 531 кодонында сериннің лейцинге аминқышқылдық алмасуы бар Ser531Leu мутациясының басымдылығын көрсетті. Біздің жұмысымызда Beijing M. tuberculosis тұқымдасы геннің 531 кодонындағы Ser531Leu мутациясымен ассоциацияланды (p<0.0001), ал LAM M. tuberculosis тұқымдасы геннің 526 кодонындағы His526Leu мутациясымен байланысты (p<0.0001) болды. Drug-resistant tuberculosis remains one of major health problems in Healthcare system of Kazakhstan. Kazakhstan is one of countries with high quantity of multidrug-resistant tuberculosis chracterized by resistance to the main antibiotics used to cure tuberculosis – rifampicin and izoniazid. Aim of this study is to detect biodiversity and characterize mutations in rpoB gene of rifampicin-resistant M. tuberculosis clinical isolates from Kazakhstan. In this work, 115 M. tuberculosis clinical isolates resistant to the first line antibiotic – rifampicin were gathered from patients with pulmonary tuberculosis. Genotyping of 115 rifampicin-resistant isolates was performed by MIRU-VNTR method using 15 loci. DNA sequencing of RRDR region of rpoB gene of 115 rifampicin-resistant isolates of M. tuberculosis was done on ABI 3730 DNA Analyzer (Applied Biosystems). According to the MIRU-VNTR genotyping results, 86,1% isolates among the resistant samples were isolates of Beijing M. tuberculosis family. Analysis of nucleotide sequences of all resistant to rifampicin isolates showed prevalence of mutations at 531 codon of rpoB gene with the amino acid substitution of serine to leucine Ser531Leu in 82.6% of cases. In our study Beijing M. tuberculosis family was associated with the Ser531Leu mutation at 531 codon of the gene (p<0.0001), LAM M. tuberculosis family revealed association with the His526Leu mutation at 526 codon of the gene (p<0.0001)
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Gautam, Praveen B., Ashwini Mishra, and Santosh Kumar. "Prevalence of rifampicin resistant mycobacterium tuberculosis and associated factors among presumptive tuberculosis patients in eastern Uttar Pradesh: a cross sectional study." International Journal Of Community Medicine And Public Health 5, no. 6 (May 22, 2018): 2271. http://dx.doi.org/10.18203/2394-6040.ijcmph20182039.
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Background: Drug resistant tuberculosis threatens global TB control and is a major public health problem in several countries and India has the highest tuberculosis in the world. The rifampicin resistance is a good predictor of multidrug resistant tuberculosis. The aim of this study was to determine the prevalence of rifampicin resistance M. tuberculosis and associated factor among presumptive tuberculosis patients in eastern Uttar Pradesh.Methods: A cross-sectional study was conducted from October 2016 to September 2017. Detection of M. tuberculosis and resistance to rifampicin was performed using Gene Xpert MTB/RIF assay. Data was collected using pre-structured questionnaire by face to face interview. The chi-square test was used to assess the statistical significance of each ratio, p<0.05 was considered significant.Results: Out of 510 patients, Mycobacterium tuberculosis was detected in 168 (32.9%). Out of these 168 patients, the prevalence of rifampicin resistance tuberculosis was 44 (26.1%). It was higher among male 38 (30.6%) than female 6 (13.6%). Regarding age distribution, maximum numbers of rifampicin resistance patients were in the age group of 20-40 years 36.7%. The prevalence of rifampicin resistance was 36 (27.6%) and 8 (21.0%) in pulmonary and extra-pulmonary respectively. Out of 44 rifampicin resistant cases, 39 (37.8%) were previously treated and 5 (7.6%) cases were treatment naïve patients. In this study, among presumptive DRTB cases, new 2 (11.7%), relapse 13 (39.3%), failure 23 (46.0%), loss to follow-up 1 (10.0%) and MDR contact 1 (20.0%) respectively were rifampicin resistant and one HIV seropositive patient was found to be rifampicin resistant.Conclusions: Previously treated cases were significantly associated with rifampicin resistance tuberculosis. The Gene Xpert is a good equipment for rapid detection and management of drug resistant tuberculosis for both pulmonary as well as extra-pulmonary tuberculosis.
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Sanzhakov, M. A., O. M. Ipatova, T. I. Torkhovskaya, E. G. Tikhonova, N. V. Medvedeva, T. S. Zakharova, and V. N. Prozorovskiy. "Increase of antituberculosis efficiency of rifampicin embedded into phospholipid nanoparticles with sodium oleate." Biomeditsinskaya Khimiya 64, no. 6 (2018): 505–10. http://dx.doi.org/10.18097/pbmc20186406505.
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The formulation of the antituberculosis drug rifampicin embedded into 20-30 nm nanoparticles from soy phosphatidylcholine and sodium oleate, is characterized by greater bioavailability as compared with free drug substance. In this study higher antituberculosis activity of this formulation was shown. Rifampicin in nanoparticles demonstrated more effective inhibition of M. tuberculosis H37Rv growth: minimal inhibiting concentration (MIC) was twice smaller than for free rifampicin. Administration of this preparation to mice with tuberculosis induced by M. tuberculosis Erdman revealed that after 6 weeks of oral administration the CUF value in lung was 22 times smaller for rifampicin in nanoparticles than for free drug (1.7 un. vs. 37.4 un.). The LD50 value in mice was two fold higher for rifampicin in nanoformulation.
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Notopuro, P. B., J. Nugraha, and H. Notopuro. "DETEKSI MOLEKUL MUTASI GEN rpoB MYCOBACTERIUM TUBERCULO SIS PADA DAHAK DENGAN POL YME RASE CHAIN REACTION DAN SINGLE STRAND CONFORMATION POLYMORPHISM." INDONESIAN JOURNAL OF CLINICAL PATHOLOGY AND MEDICAL LABORATORY 16, no. 2 (March 16, 2018): 81. http://dx.doi.org/10.24293/ijcpml.v16i2.973.
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Tuberculosis is a chronic infectious disease which is found in the developing as well as the developed country. This disease is oneof the community health problems which become the priority programs in the national as well as international health. In the lasttwo decades, they can be found in the emergency tuberculosis problems that is related with the Multi Drug Resistance (MDR) Strain.The detection of rifampicin resistance in M. tuberculosis infection can help clinical laboratory to find the MDR strain. Related to thisproblem the proportional culture method is still the gold standard for rifampicin resistance detection for M. tuberculosis infection. Butthis method needs 4−6 weeks to obtain the result, while its sensitivity is not very high. The development of the molecular detection forM. tuberculosis rifampicin resistance in a direct clinical specimen such as sputum, cerebrospinalfLuid, etc. will give an improvement inthe diagnosis, because it has an accurate, fast, sensitive and a specific result. Isolates from twenty six of M. tuberculosis derived fromthe sputum of tuberculosis patients that have failed the tuberculosis treatment, were examined with the proportional culture method.In this study PCR-SSCP were used for the molecular detection of rifampicin resistancy using direct sputum samples. The proportionalculture method was used as a gold standard for the rifampicin resistance detection. A set of primers was directed to conserve the regionof rpoB gene of M. tubercuLosis. This RNA polymerase gene was encondes?, which is bound on rifampicin. A 157-bp fragment wasamplified by PCR and analyzed by SSCP technique. The sensitivity of PCR-SSCP is 80% (high), its specificity is 95.2% (very high), thepositive predictive value is 80% and the negative predictive value is 95.2%. Statistically there were no significant difference between theresult of PCR-SSCP and the proportional culture method. Based on the study result, the molecular detection technique for rifampicinresistance on M. tuberculosis infection can be used as the screening device /means for Multi Drug Resistance Tuberculosis (MDR-TB),while the clinician waits the culure result.
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N. Djide, Nana Juniarti, M. Natsir Djide, Muhammad Nur Amir, and Sartini Sartini. "Aktivitas Antibakteri Ekstrak Kelopak Bunga Rosella Terenkapsulasi Maltodekstrin dan Sinergitasnya dengan Isoniazida dan Rifampisin Terhadap Mycobacterium tuberculosis H37rv." Jurnal Farmasi Galenika (Galenika Journal of Pharmacy) 5, no. 2 (October 15, 2019): 117. http://dx.doi.org/10.22487/j24428744.0.v0.i0.12946.
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First-line drugs (Isoniazid and Rifampicin) for the treatment of tuberculosis are known to have experienced resistance to Mycobacterium tuberculosis. The aim of this study was to determine the minimum inhibitory concentration (MIC) of maltodextrin encapsulated rosella calyx extract and its ability to provide a synergistic effect with Isoniazid (INH) and Rifampicin (RIF) on M. tuberculosis R37rv. Rosella calyxs were macerated using 50% ethanol and encapsulated using maltodextrin. Antibacterial activity was carried out by determining MIC value using Microscopic Observation and Direct Susceptibility (MODS) method. The synergistic effect was carried out by calculating the Fractional Inhibition Concentration Index (FICI). The results showed that this extract was able to inhibit M. tuberculosis H37rv with MIC of 10 mg/ ml. The FICI value of the combination of extract with INH and rifampicin was obtained 1.25. This showed that rosella calyx extract is not synergistic with INH and rifampicin, might be due to rosella calyx extract has an antibacterial effect with a different mechanism with INH and Rifampicin.
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N. Djide, Nana Juniarti, M. Natsir Djide, Muhammad Nur Amir, and Sartini Sartini. "Aktivitas Antibakteri Ekstrak Kelopak Bunga Rosella Terenkapsulasi Maltodekstrin dan Sinergitasnya dengan Isoniazida dan Rifampisin Terhadap Mycobacterium tuberculosis H37rv." Jurnal Farmasi Galenika (Galenika Journal of Pharmacy) (e-Journal) 5, no. 2 (October 15, 2019): 117–23. http://dx.doi.org/10.22487/j24428744.2019.v5.i2.12946.
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First-line drugs (Isoniazid and Rifampicin) for the treatment of tuberculosis are known to have experienced resistance to Mycobacterium tuberculosis. The aim of this study was to determine the minimum inhibitory concentration (MIC) of maltodextrin encapsulated rosella calyx extract and its ability to provide a synergistic effect with Isoniazid (INH) and Rifampicin (RIF) on M. tuberculosis R37rv. Rosella calyxs were macerated using 50% ethanol and encapsulated using maltodextrin. Antibacterial activity was carried out by determining MIC value using Microscopic Observation and Direct Susceptibility (MODS) method. The synergistic effect was carried out by calculating the Fractional Inhibition Concentration Index (FICI). The results showed that this extract was able to inhibit M. tuberculosis H37rv with MIC of 10 mg/ ml. The FICI value of the combination of extract with INH and rifampicin was obtained 1.25. This showed that rosella calyx extract is not synergistic with INH and rifampicin, might be due to rosella calyx extract has an antibacterial effect with a different mechanism with INH and Rifampicin.
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Mohiuddin, Md, and J. Ashraful Haq. "First Line Anti-Tubercular Drug Resistance Pattern of Mycobacterium Tuberculosis Isolated From Specialized Hospitals of Dhaka City." Ibrahim Medical College Journal 8, no. 2 (February 4, 2016): 41–46. http://dx.doi.org/10.3329/imcj.v8i2.26677.
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The present study was undertaken to determine the drug resistance pattern of M. tuberculosis isolated from 225 pulmonary and 45 extrapulmonary tuberculosis cases. The samples were cultured on Lowenstein Jensen (L-J) media for isolation of M. tuberculosis. Drug resistance to first line anti tubercular drugsnamely isoniazid (INH), rifampicin (RIF), Ethambutol (ETH) and streptomycin (SM) were determined by indirect proportion method. The overall drug resistance of M. tuberculosis was 53.6% to any of the first line anti tubercular drugs. Rate of multi drug resistant tuberculosis (MDR-TB) among the untreated cases was 4.2%, while it was 36.0% in previously treated cases. It was found that 83.3% rifampicin resistant M. tuberculosis was cross resistant to one or more of other first line anti-tubercular drugs, while cross resistance of INH, ETH and SM resistant isolates was much low. The present study revealed that high level of drug resistance exists to individual anti tubercular drugs and MDR-TB is an emerging problem, particularly in treated cases. Rifampicin resistance could be used as a surrogate marker for drug resistance to other first line anti tubercular drugs.Ibrahim Med. Coll. J. 2014; 8(2): 41-46
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Kashyap, Bineeta, Nisha Goyal, Puneeta Hyanki, NP Singh, and Ashwani Khanna. "Cartridge-based nucleic acid amplification test: a novel rapid diagnostic tool to study the burden of tuberculosis from a tertiary care hospital." Tropical Doctor 49, no. 4 (July 10, 2019): 274–81. http://dx.doi.org/10.1177/0049475519859958.
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Despite efforts to limit the morbidity and mortality from tuberculosis (TB), it continues to be an important cause of death. There is an urgent need for a diagnostic test that accurately and quickly diagnoses TB, especially if it is also a near-point-of-care test. The GeneXpert polymerase chain reaction test (known in India as CBNAAT [cartridge-based nucleic acid amplification test] and is capable of diagnosing TB and rifampicin resistance within 2 h) is a promising tool. The duration of our study was two years and was carried out in the DOTS centre of a tertiary care hospital in India. A total of 5449 samples were processed using CBNAAT. Of the total samples tested, 2068 were extra-pulmonary. The following information was collected: number of extra-pulmonary samples processed; number of Mycobacterium tuberculosis ( M. tuberculosis)-positive samples; patterns of rifampicin sensitivity; number of people living with HIV (PLHIV); and number of children. Of the samples, 62.1% were from suspected pulmonary TB patients. Out of the total samples tested using CBNAAT, 21.8% were positive for M. tuberculosis. Rifampicin resistance was seen in 9.2%, 8.5% and 10.3% of the total, pulmonary and extra-pulmonary samples, respectively, in M. tuberculosis-positive samples. Overall, 36.9% samples were from the paediatric population and 5.7% belonged to PLHIV. Rifampicin resistance was seen in 8.8% and 8.3% of the M. tuberculosis-positive paediatric and PLHIV samples, respectively.
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Zahoor, Danish, Shameem Wani, and Zaffer Nowshad Wani. "Evaluation of cepheid GeneXpert MTB/RIF assay for Mycobacterium tuberculosis detection and rifampicin resistance in clinical specimen." International Journal of Research in Medical Sciences 7, no. 6 (May 29, 2019): 2121. http://dx.doi.org/10.18203/2320-6012.ijrms20192484.
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Background: Timely diagnosis and treatment of tuberculosis is important to treat the disease and to reduce transmission. The WHO recommends using GeneXpert MTB in developing, high-burden countries. A study was conducted to evaluate the performance of Xpert assay for the detection of M. tuberculosis and rifampicin resistance in clinical specimen.Methods: About 615 consecutive samples were simultaneously subjected to culture and phenotypic drug susceptibility test for M. tuberculosis and analysis by GeneXpert assay. Confirmed Mycobacterium tuberculosis in a positive culture was used as a reference standard for TB diagnosis.Results: The assay achieved a sensitivity of 96.75% (268/277) and 76.47% (26/34) for smear positive and smear negative pulmonary specimen respectively. In extrapulmonary specimen, the sensitivity was 50% (1/2) and 42.8% (3/7) for smear positive and smear negative specimen respectively. An additional 48 M. tuberculosis were detected by Xpert assay which were smear and culture negative. The Xpert assay identified 100% of the phenotypic rifampicin susceptible isolates and 74.19% of the phenotypic rifampicin resistant isolates. Discordant results were seen in 8 (2.76%) isolates. 6 of these isolates were confirmed to be rifampicin resistant by the reference lab.Conclusions: Present study indicates that Xpert MTB/RIF assay is an effective and rapid tool for the rapid diagnosis of Mycobacterium tuberculosis. The sensitivity is comparable to culture in smear positive specimen but less sensitive than culture for smear negative specimen. In cases with high index of suspicion or discordance for rifampicin results, confirmation should be done by other methods due to false negative results on Xpert assay.
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Laskar, Nourjahan, Md Akram Hossain, Jannatul Fardows, and Mominur Rahman. "GeneXpert MTB/RIF Assay for Rapid Identification of Mycobacterium Tuberculosis and Rifampicin Resistance Directly from Sputum Sample." Journal of Enam Medical College 7, no. 2 (June 4, 2017): 86–89. http://dx.doi.org/10.3329/jemc.v7i2.32653.
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Background: The World Health Organization has endorsed the use of molecular methods for the detection of tuberculosis (TB) and drug resistant TB as a rapid method. In Bangladesh, the Xpert MTB/RIF assay has been implemented into reference laboratories for diagnosis of TB and also MDR TB.Objective: Drug resistant tuberculosis has long been a common problem prevailing in our country. The present study focused on the rapid identification of Mycobacterium tuberculosis as well as drug resistance.Materials and Methods: Sputum samples from a total of 107 cases, assumed as multi-drug resistance tuberculosis, were studied through GeneXpert assay.Results: Out of 107 cases, 91 (85.05%) were detected having M. tuberculosis ? 64 (59.81%) were rifampicin sensitive and 27 (25.23%) were rifampicin resistant. The sensitivity and specificity of the GeneXpert are 87.64% and 75% respectively.Conclusion: GeneXpert assay can be considered for the rapid diagnosis of drug resistant tuberculosis.J Enam Med Col 2017; 7(2): 86-89
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Huey, Lee Lih, Chua Wei Chuan, Zeti Norfidiyati Ayub Salmuna, Norazmi Mohd Nor, Alwi Muhd Besari and Chan, and Yean Yean. "A novel two-codon deletion at codon 508 and 509 of the rpoB gene in pulmonary tuberculosis and HIV co-infection: A case report." Pakistan Journal of Medical and Health Sciences 16, no. 7 (July 30, 2022): 201–2. http://dx.doi.org/10.53350/pjmhs22167201.
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We report a case of pulmonary Mycobacterium tuberculosis infection with a resistance to rifampicin (RIF) in a 39-year-old man who was co-infected with Human Immunodeficiency Virus (HIV) and hepatitis C. Phenotypic and genotypic test were performed to diagnose the patient. Sequencing analysis revealed novel two-codon deletion at codon 508 and 509 of the rpoB gene in M. tuberculosis. Keywords: Co-infection, Mycobacterium tuberculosis, rpoB gene, rifampicin-resistant.
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Tesic, Jelena, Zorica Lepsanovic, and Veljko Mirovic. "Evaluation of usefulness of single-strand conformation polymorphism method for rapid detection of rifampicin-resistant mycobacterium tuberculosis." Vojnosanitetski pregled 61, no. 2 (2004): 169–72. http://dx.doi.org/10.2298/vsp0402169t.
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The aim was to evaluate the Single-Strand Conformation Polymorphism method as a potential tool for rapid detection of rifampicin-resistant strains by the use of 39 rifampicin-resistant Mycobacterium tuberculosis strains isolated in Serbia. SSCP analysis on acrylamide gel detected 56.4% of the rifampicin-resistant M. tuberculosis strains and showed the inability to detect one of the most frequent mutations, TCG?TTG mutation in codon 531 of the rpoB gene, which was shown by automated sequencing.
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Riwu, Audrey Gracelia, Jusak Nugaraha, and Yoes Prijatna Dachlan. "DIFFERENCES OF INTERLEUKIN-18 AND INTERLEUKIN-10 LEVELS IN RIFAMPICIN RESISTANT AND RIFAMPICIN SENSITIVE PULMONARY TUBERCULOSIS PATIENTS IN DR. SOETOMO HOSPITAL SURABAYA." Indonesian Journal of Tropical and Infectious Disease 8, no. 2 (July 31, 2020): 116. http://dx.doi.org/10.20473/ijtid.v8i2.10959.
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Rifampicin is an anti-tuberculosis drug which has an efficient antimicrobial effect and the basis of a short-term treatment regimen for tuberculosis (TB) patients. Rifampicin plays an important role against the growth and slow metabolism of Bacilli M. tuberculosis. Resistance to rifampicin causes the duration of tuberculosis treatment to be longer. Interleukin-18 (IL-18) is a pro-inflammatory cytokine which plays a role in controlling the growth of M. tuberculosis through its ability to induce IFN-γ, while Interleukin-10 (IL-10) is an anti-inflammatory cytokine which plays a role in limiting tissue damage due to the inflammatory process and maintain tissue homeostasis. IL-18 and IL-10 has an important role in explaining the different degrees of inflammation in rifampicin resistant (RR) and rifampicin sensitive (RS) pulmonary tuberculosis patients. The purpose of this study is to determine different levels of IL-18 and IL-10 in new TB patients with RR and RS. This study was a retrospective cohort study with a cross-sectional design carried out from August-November 2018 in the TB-DOTS/MDR clinic at Dr. Soetomo Hospital, Surabaya. 50 research subjects were examined and grouped into two groups, namely pulmonary TB with RR (n = 25) and pulmonary TB with RS (n = 25) based on GeneXpert examination and anti-tuberculosis drug therapy ≤ 1 month. IL-18 and IL-10 were measured using the ELISA Method. Differences in IL-18 and IL-10 levels between groups were analyzed using the Mann-Whitney test. The mean level of IL-18 (pg/ml) in RR and RS pulmonary TB patients were 1273.53±749.86 and 787.96 ±589.28 respectively. The mean level of IL-10 (pg/ml) in RR and RS pulmonary TB patients were 125.25±118.32 and 128.81±135.77 repectively. The mean level of IL-18 in RR and RS pulmonary TB patients were found to have a significant difference, while the mean level of IL-10 did not have a significant difference. Keywords: Interleukin-18, Interleukin-10, Tuberculosis, Rifampicin Resistant, Rifampicin Sensitive
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Jiang, Haiqin, Lemuel Tsang, Hongsheng Wang, and Changhong Liu. "IFI44L as a Forward Regulator Enhancing Host Antituberculosis Responses." Journal of Immunology Research 2021 (October 20, 2021): 1–12. http://dx.doi.org/10.1155/2021/5599408.
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Interferon-induced protein 44-like (IFI44L) gene is a type I interferon-stimulated gene (ISG) that plays a critical role in antiviral activity and constitutes a promising diagnostic marker. However, its precise role and function in tuberculosis have not been unveiled. This study showed that IFI44L acts as an antimicrobial target and positive modulator in human macrophages. Knockdown of IFI44L led to increased Mycobacterium tuberculosis intracellular survival. Moreover, IFI44L was significantly upregulated, and it restricted the intracellular survival of M. tuberculosis H37Rv strains at 72 h after rifampicin treatment. Individuals with cutaneous tuberculosis (CTB) were found to have significantly higher IFI44L expression after 6 months of rifampicin therapy than after only 1 month. These results demonstrated that IFI44L induced positive regulation and clearance of M. tuberculosis from human macrophages. This antimicrobial activity of IFI44L makes it a possible target for therapeutic applications against M. tuberculosis.
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Zeng, Sheng, Dong Yang, Céline Rens, and Véronique Fontaine. "The Mycobacterium bovis BCG GroEL1 Contributes to Isoniazid Tolerance in a Dormant-Like State Model." Microorganisms 11, no. 2 (January 21, 2023): 286. http://dx.doi.org/10.3390/microorganisms11020286.
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Due to the Mycobacterium tuberculosis complex, including M. tuberculosis and M. bovis, tuberculosis still causes 1.6 million deaths per year. Therefore, efforts to improve tuberculosis treatment are necessary. We previously showed that the GroEL1 protein is involved in antibiotic intrinsic resistance. Indeed, the M. bovis BCG cpn60.1 gene (encoding GroEL1)-disrupted strain (Δcpn60.1) exhibits higher rifampicin and vancomycin susceptibility due to defective cell wall integrity. Here, we show that during hypoxia-triggered growth stasis, in the Wayne dormancy model, the mutant exhibited comparable rifampicin and ethionamide susceptibility but higher isoniazid susceptibility compared to the wild-type strain. Although the Δcpn60.1 strain showed compromised induction of the DosR regulon, growth stasis was achieved, but an ATP burst and a higher reactive oxygen species (ROS) production were observed in the isoniazid-treated Δcpn60.1 strain. GroEL1 could contribute to INH tolerance by reducing ROS.
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Sabaté, M. Betriu, A. González-Rodríguez, A. Guàrdia, E. Pérez-Macho, A. Alvarez Pedrero, S. Acebillo, J. A. Monreal, D. Palao Vidal, and J. Labad. "Pharmacokinetic interactions of psychotropic medications in patients with schizophrenia suffering from atypical mycobacterial infections." European Psychiatry 64, S1 (April 2021): S649—S650. http://dx.doi.org/10.1192/j.eurpsy.2021.1725.
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IntroductionMycobacterium kansasii is a nontuberculous mycobacterium that causes infection associated with past or current tuberculosis disease. Clinical syndromes and radiological findings are mostly indistinguishable from that of Mycobacterium tuberculosis, thus requiring microbiological confirmation.ObjectivesWe report a case of a 44-year-old man diagnosed with schizophrenia and Mycobacterium kansasii infection.MethodsCase report and non-systematic narrative review from PubMed.ResultsCase report: Patient with schizophrenia who was admitted at the inpatient unit presenting psychotic exacerbation with high levels of excitement. Risperidone 6 mg/day and valproate 500 mg/day were initiated. He was also diagnosed with a M. kansasii lung infection, with radiological findings of past tuberculosis disease. Before the microbiological confirmation, it was necessary to start rifampicin, requiring an increase in doses of both psychotropic drugs. Review: (1)Comorbidity of mycobacterial infections and schizophrenia. Several studies have shown that people with severe mental illness have higher rates of tuberculosis compared with the general population. Although the relationship between tuberculosis and M. Kansasii infection is known, few literature is available with regard to the association of M. Kansasii and schizophrenia. (2)Interactions between antipsychotics and mood stabilizers with rifampicin. Rifampicin is mainly metabolized by CYP3A4 and transported by P-glycoprotein. Add-on with rifampicin have been reported to reduce clozapine and olanzapine plasma levels (despite both are metabolized by CYP1A2), reduce haloperidol and risperidone levels (possible role of P-glycoprotein in this interaction), as well as for valproate.ConclusionsTreatment of comorbid infections in people with schizophrenia remains a challenge. Antibiotics used to treat mycobacterial infections can modify the pharmacokinetic of psychotropic drugs.DisclosureNo significant relationships.
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Rahman, Fahmida, Sadia Sharmin, Md Mustafa Kamal, and Md Ruhul Amin Miah. "Drug resistance pattern of M. tuberculosis in category II treatment failure pulmonary tuberculosis patients." Ibrahim Medical College Journal 7, no. 1 (January 20, 2014): 9–11. http://dx.doi.org/10.3329/imcj.v7i1.17741.
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This study was designed to determine the extent of drug resistance of M. tuberculosis (MTB) isolated from category II treatment failure pulmonary tuberculosis (PTB) patients. A total of 100 Ziehl-Neelsen (Z-N) smear positive category II failure PTB patients were included in this study. Sputum culture was done in Lowenstein-Jensen (L-J) media. Conventional proportion method on Lowenstein-Jensen (L-J) media was used to determine the drug susceptibility of M. tuberculosis to isoniazid (INH), rifampicin (RMP), ofloxacin (OFX) and kanamycin (KA). Out of 100 sputum samples, a total of 87 samples were positive by culture. Drug susceptibility test (DST) revealed that 82 (94.25%) isolates were resistant to one or more anti -TB drugs. Resistance to isoniazide (INH), rifampicin (RMP), ofloxacin (OFX) and kanamycin (KA) was 94.25%, 82.75%, 29.90% and 3.45% respectively. Among these isolates, 79.31% and 3.45% isolates were multi-drug resistant (MDR) and extended drug resistant (XDR) M. tuberculosis respectively. High rate of anti-tubercular drug resistance was observed among the category II treatment failure TB patients.DOI: http://dx.doi.org/10.3329/imcj.v7i1.17741 Ibrahim Med. Coll. J. 2013; 7(1): 9-11
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Murthy, Neetha S., Sanjeev Nair, PK Ramya, Renu Susan George, B. Ravikrishnan, M. Sunil Kumar, Sairu Philip, PS Rakesh, and Shibu Balakrishnan. "Drug-resistant Tuberculosis: First Line Drug Resistance Pattern among Mycobacterium Tuberculosis Strains Isolated from a Reference Laboratory in Kerala State, India." Journal of Pure and Applied Microbiology 15, no. 4 (October 4, 2021): 1882–91. http://dx.doi.org/10.22207/jpam.15.4.09.
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Resistance to antimycobacterial agents consistently remains a major obstacle to end TB in India. Geographical prevalence data regarding drug-resistant evolutionary genetics of M. tuberculosis (MTB) remains sparse in India. Our objective was to determine the genotypic drug resistance mutation pattern for Rifampicin and Isoniazid of MTB isolates to gain an understanding of the prevailing molecular epidemiology of drug-resistant tuberculosis. In this study 2528 M. tuberculosis DNA isolates from presumptive DRTB suspects received at the nodal TB reference laboratory in Kerala were tested for Rifampicin and Isoniazid resistance by sequence-based diagnostic Line Probe assay (LPA). Geographical prevalence and associations of rpoB, katG, inhA resistance codons was analyzed from January 2019 to March 2020. Among the 2528 DNA samples subjected for Rifampicin and Isoniazid resistance determination by LPA, 146 (5.8%) isolates were resistant to both drugs. Isoniazid mono-resistance was found in 164 (6.5%) and Rifampicin mono-resistance in 38 (1.5%) isolates. The most frequent rpoB mutation was S531L (60.32%) followed by S531W/L533P mutations seen in 8.15% of the isolates. S315T1 KatG mutation was seen in 97.33% of Isoniazid resistant isolates. 84.68% isolates with rpoB S531L mutation were found to be multidrug-resistant. 82.9% of isolates with rpoB S531L mutation showed katG S315T1 mutation. Mono isoniazid-resistant isolates were significantly higher compared to mono rifampicin-resistant isolates among the DNA isolates studied in our region. The molecular epidemiological pattern most frequently associated with multidrug resistance was rpoB S531L which was significantly associated with the co-presence of S315T1 mutation.
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Garg, Saurabh, Rakesh Kumar, Dennis Kunimoto, and Gina R. Rayat. "Anti-Mycobacterial Activity of Flavonoid and Pyrimidine Compounds." Molecules 27, no. 19 (October 9, 2022): 6714. http://dx.doi.org/10.3390/molecules27196714.
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We evaluated the anti-mycobacterial effect of a flavonoid 5,7-dihydroxy-2-(4-hydroxyphenyl) 4H-chromen-4-one (1) and two pyrimidines, 4-hydroxy-2-dimethylamino-5-nitroso-6-aminopyrimidine (2) and 2-chloro-5-n-nonylpyrimidine (3) in vitro against Mycobacterium tuberculosis (M. tuberculosis, H37Ra) and Mycobacterium avium (M. avium), using a Microplate Alamar Blue Assay (MABA). The effects of the compounds 1–3 in combination with first- and second-line anti-TB drugs isoniazid, rifampicin, cycloserine, and clarithromycin on the growth of M. tuberculosis and M. avium were also evaluated in in vitro assays. As a single agent, compounds 1 and 2 exhibited modest activity while compound 3 was the most effective against M. tuberculosis and M. avium. When compounds 1–3 were evaluated at lower than 50% of their inhibitory concentrations in a two-drug combination with isoniazid or rifampicin, they showed additive to synergistic interactions. This inhibitory effect was improved when each of the three compounds was tested together in a three-drug combination with two of the first-line anti-TB drugs. Compounds 1–3 also demonstrated strong synergistic interaction in combination with cycloserine and clarithromycin in inhibiting the growth of M. tuberculosis and M. avium, respectively. This study demonstrated that compounds 1–3 have potential to be developed as effective anti-TB agents with combined use.
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Coban, Ahmet Yilmaz. "A novel agar base medium for drug susceptibility testing of Mycobacterium tuberculosis isolates." Future Microbiology 16, no. 13 (September 2021): 949–53. http://dx.doi.org/10.2217/fmb-2021-0032.
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Aim: In this study, it was aimed to evaluate AYC.2.2 agar for susceptibility testing of Mycobacterium tuberculosis clinical isolates against first-line drugs. Materials & methods: In the present study, 208 M. tuberculosis clinical isolates were tested on AYC.2.2 agar, which was previously validated for the first-line drugs isoniazid, rifampicin, streptomycin and ethambutol. Results: Specificity, sensitivity, positive predictive value, negative predictive value and agreement for isoniazid–rifampicin–ethambutol–streptomycin were 100–100–97.2–99.3%, 94.8–94.8–79.3–94.3%, 100–100–82.1–98.03%, 97.03–98.03–96.7–98.08%, 98.07–98.5–94.7-98.07%, respectively. Conclusion: Results had shown that the newly developed AYC.2.2 agar promises as an alternative medium that can be used to perform susceptibility testing of M. tuberculosis isolates. However, further multicenter studies are needed to be used in routine mycobacteriology laboratories.
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Zhurilo, A. A., A. I. Barbova, Yu A. Cherednik, P. S. Trofimova, S. V. Mironchenko, O. V. Pavlova, A. V. Chernov, and L. M. Sladkova. "COMPARISON OF GENEXPERT MTB/RIF AND GENOTYPE SYSTEMS WITH MTBDRPLUS STRIPS FOR DETECTION OF MUTATIONS THAT ARE ASSOCIATED WITH M. TUBERCULOSIS RESISTANCE TO RIFAMPICIN IN TUBERCULOSIS." Ukrainian Pulmonology Journal 30, no. 4 (2022): 34–41. http://dx.doi.org/10.31215/2306-4927-2022-30-4-34-41.
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COMPARISON OF GENEXPERT MTB/RIF AND GENOTYPE SYSTEMS WITH MTBDRPLUS STRIPS FOR DETECTION OF MUTATIONS, ASSOCIATED WITH M. TUBERCULOSIS RESISTANCE TO RIFAMPICIN IN TUBERCULOSIS A. A. Zhurilo, A. I. Barbova, Yu. A. Cherednik, P. S. Trofi mova, S. V. Mironchenko, O. V. Pavlova, A. V. Chernov, L. M. Sladkova Summary The aim was to analyze the level of compliance of two molecular genetic methods GeneXpert MTB/RIF and GenoTypeDRplus in determining the drug resistance of M. tuberculosis to rifampicin when detecting mutations in the RRDR region associated with drug resistance. Object and methods. We studied strains of M. tuberculosis with resistance to rifampicin, which was detected by any of the studied molecular genetic methods. 96 sputum samples were taken. Sputum smears were examined for the presence of acid-fast bacteria by microscopy after staining with the Ziehl-Neelsen method. The material was inoculated into Middlebrook 7H9 broth and on Lowenstein-Jensen medium. The liquid medium was incubated in the BACTEC MGIT system. An immunochromatographic test was used to identify the strains. The drug susceptibility test of M. tuberculosis to rifampicin was performed using the BACTEC MGIT system. The GeneXpert MTB/RIF test was performed according to the manufacturer’s instructions. The GenoTypeDRplus assay was performed on decontaminated and concentrated sputum samples. The process was carried out in three stages: DNA extraction from the processed sputum sample; amplification of the RRDR region by PCR; hybridization of the PCR product to specific oligonucleotide probes immobilized on the test strip. For sequencing, M. tuberculosis DNA isolation was performed using the QIAamp® DNA Mini Kit. The DNA concentration was measured on a Denovix Quantus spectrophotometer. Targeted panel amplification was performed using the Deeplex Myc-TB kit. Amplicon purification was performed using Agencourt AMPure XP magnetic beads. Quantitative analysis of the purified amplification products was performed using a Qubit fluorometer. The M. tuberculosis DNA library was prepared for sample sequencing using the Nextera XT DNA library preparation kit. The library used 5.0 μl of input DNA at a concentration of 0.2 ng/μl. Sequencing was performed on MiSeq equipment with the library normalization and denaturation protocol. Results and discussion. The GeneXpert MTB/RIF and MTBDRplus systems target the same 81 bp rifampicin resistance domain. (RRDR) subunits of bacterial RNA polymerase (rpoB) for mutation detection using DNA probes, i.e. there is a correspondence of probes to each other and an expected similarity of probe binding. We analyzed all sputum samples using GeneXpert MTB/RIF and GenoType MTBDRplus and phenotypic BACCTEC MGIT methods. The level of agreement between two molecular genetic methods for the detection of rifampicin-associated mutations in the RRDR region has been established. The RRDR 81bp region of the rpoB gene of mismatched cases was studied by sequencing. GeneXpert MTB/RIF and GenoType DRplus matched the phenotypic method in 92.7% and 89.6% of cases of M. tuberculosis resistance, respectively. Complete agreement between the results of GeneXpert MTB/RIF and GenoTypeMTBDRplus was observed in 92.7% of cases. GeneXpert MTB/RIF and GenoType DRplus showed a similar pattern of binding failure of wild type probes (WT-probes) when scanning the 81 bp region (RRDRdomain), which leads to stability diagnostics through probe failure software. Sequencing of the RRDR region of “mismatched” strains showed that GeneXpert probes detected seven “mismatched” cases correctly, and GenoTypeDRplus was erroneous in all cases. GeneXpert has demonstrated greater accuracy in R-resistance detection for mismatched isolates compared to GenoTypeDRplus. GeneXpert MTB/RIF has a number of other benefits over GenoTypeDRplus. GeneXpert MTB/RIF is relatively easier to implement, biosafety requirements are minimal, study times are shorter, and the study process is more automated, resulting in less human error and more reproducible results. Given these facts and the results of GeneXpert MTB/RIF found in the study, it is recommended that GeneXpert MTB/RIF be used to detect MDR-TB. Conclusions. Sequencing of the 81bp RRDR region of mismatched M. tuberculosis strains showed that GeneXpert MTB/RIF performed more accurately than GenoTypeDRplus in detecting mutations associated with rifampicin resistance. GeneXpert MTB/RIF is relatively easier to perform, biosafety requirements are minimal, time to study is shorter, and the study process is more automated, resulting in less human error and greater reproducibility of results, so it is reasonable to use it for the detection of multidrug-resistant tuberculosis. The Deeplex Myc-TB analytical solution delivers a wealth of insightful information from antimycobacterial drug resistance markers and speeds up data analysis with easy-to-use software. Key words: tuberculosis, Mycobacterium tuberculosis, drug resistance, GeneXpert MTB/RIF and GenoTypeDRplus molecular genetic systems, BACTEC MGIT phenotypic system, sequencing. Ukr. Pulmonol. J
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Kumar Sah, Ranjit, Dwij Raj Bhatta, Gokarna Raj Ghimire, Bhuwaneswor Prasad Kandel, Bishnu Raj Tiwari, and Jeevan Bahadur Sherchand. "Evaluation of Nitrate Reductase Assay For Rapid Detection of Drug Resistant Tuberculosis." Journal of Health and Allied Sciences 3, no. 1 (November 24, 2019): 44–46. http://dx.doi.org/10.37107/jhas.53.
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Emergence of multidrug-resistant tuberculosis (MDR-TB) urgently demands for simple, rapid and inexpensive methods of its detection for the effective treatment of drug resistant tuberculosis, particularly in low-income countries. This prospective study was carried out at National Tuberculosis Reference Laboratory and South Asian Association of Regional Cooperation (SAARC) Tuberculosis and HIV/AIDS Centre, Thimi, Bhaktapur, Nepal, from November 2009 to May 2010 to evaluate nitrate reductase assay (NRA) efficacy for rapid determination of streptomycin, isoniazid, rifampicin and ethambutol susceptibility of Mycobacterium tuberculosis strains. A total of 113 clinical isolates of M. tuberculosis were tested for four first line antitubercular drugs by nitrate reductase assay and were compared with standard proportion method. Results were available in 7-14 days by NRA as compared to proportion method which generally takes 4-6 weeks. The sensitivity and specificity of NRA were 98.1% and 100% for isoniazid, 95.1% and 98.6% for rifampicin, 91.4% and 94.9% for streptomycin, and 78.6% and 97.9% for ethambutol respectively. Agreement between NRA and proportion method were 99.1%, 97.3%, 93.8%, 95.6% for isoniazid, rifampicin, streptomycin and ethambutol, respectively. NRA is easier, inexpensive and reliable method for susceptibility testing of Mycobacterum tuberculosis for isoniazid and rifampicin, the two most important drugs for the treatment of tuberculosis. The reduction in susceptibility testing time, and higher sensitivity and specificity of NRA method is of fundamental importance in detecting MDR-TB.
Key words: Drug susceptibility, MDR-TB, NRA, proportion method
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Andreevskaya, S. N., T. G. Smirnova, E. E. Larionova, I. Yu Andrievskaya, L. N. Chernousova, and A. Ergeshov. "Isoniazid-resistant Mycobacterium tuberculosis: prevalence, resistance spectrum and genetic determinants of resistance." Bulletin of Russian State Medical University, no. (1)2020 (January 12, 2020): 21–26. http://dx.doi.org/10.24075/brsmu.2020.001.
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The lack of simple, rapid diagnostic tests for isoniazid-resistant rifampicin-susceptible tuberculosis infection (Hr-TB) can result in low treatment efficacy and further amplification of drug resistance. Based on the clinical data, this study sought to estimate the prevalence of Hr-TB in the general population and characterize the phenotypic susceptibility and genetic determinants of isoniazid resistance in M. tuberculosis strains. Molecular-genetic and culture-based drug susceptibility tests were performed on M. tuberculosis isolates and M. tuberculosis DNA obtained from the patients with pulmonary TB undergoing treatment at the Central Tuberculosis Research Institute between 2011 and 2018. The tests revealed that Hr-TB accounted for 12% of all TB cases in the studied sample. Hr-TB strains were either resistant to isoniazid only (45%) or had multiple resistance to 2–6 anti-TB agents. Resistance to isoniazid was caused by mutations in the katG gene. Based on the literature analysis and our own observations, we emphasize the importance of developing simple molecular drug susceptibility tests capable of detecting simultaneous resistance to rifampicin and isoniazid and the necessity of their translation into clinical practice.
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Santos, Nathally C. de S., Regiane B. de L. Scodro, Dayane CB Leal, Silvia MT do Prado, Daniela F. Micheletti, Eloísa G. Sampiron, Giovana F. Costacurta, et al. "Determination of minimum bactericidal concentration, in single or combination drugs, against Mycobacterium tuberculosis." Future Microbiology 15, no. 2 (January 2020): 107–14. http://dx.doi.org/10.2217/fmb-2019-0050.
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Aim: To evaluate an assay to detect minimum bactericidal concentration (MBC) in Mycobacterium tuberculosis, using as single model rifampicin, isoniazid, levofloxacin (LVX) and linezolid (LNZ) and in combination. Material & methods: MBCs were carried out directly from resazurin microtiter assay plate and 3D checkerboard in M. tuberculosis H37Rv and five resistant clinical isolates. Results: The proposed MBC assay showed similar values to those determined by MGIT™, used as control. LVX and LNZ's MBC values were close to their MIC values. LNZ or LVX combined with isoniazid and rifampicin showed MBC value reduced in 63.7% of the assays. Conclusion: The proposed assay to determine MBCs of drugs can be applied to the study of new compounds with anti- M. tuberculosis activity to detect their bactericidal effect and also in laboratory routine for clinical dose adjustment of drugs according to the patient's profile.
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Sah, Ranjit Kumar, DR Bhatta, GR Ghimire, BP Kandel, BR Tiwari, and JB Sherchand. "Evaluation of Nitrate Reductase Assay for Rapid Detection of Drug Resistant Tuberculosis." SAARC Journal of Tuberculosis, Lung Diseases and HIV/AIDS 9, no. 2 (April 23, 2013): 4–8. http://dx.doi.org/10.3126/saarctb.v9i2.7971.
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Introduction: Emergence of multidrug-resistant tuberculosis (MDR-TB) urgently demands for simple, rapid and inexpensive methods of its detection for the effective treatment of drug resistant tuberculosis, particularly in low-income countries. Methodology: This prospective study was carried out at National Tuberculosis Reference Laboratory and South Asian Association of Regional Cooperation (SAARC) Tuberculosis and HIV/AIDS Centre, Thimi, Bhaktapur, Nepal, from November 2009 to May 2010 to evaluate nitrate reductase assay (NRA) efficacy for rapid determination of streptomycin, isoniazid, rifampicin and ethambutol susceptibility of Mycobacterium tuberculosis strains. Results: A total of 113 clinical isolates of M. tuberculosis were tested for four first line antitubercular drugs by nitrate reductase assay and were compared with standard proportion method. Results were available in 7-14 days by NRA as compared to proportion method which generally takes 4-6 weeks. The sensitivity and specificity of NRA were 98.1% and 100% for isoniazid, 95.1% and 98.6% for rifampicin, 91.4% and 94.9% for streptomycin, and 78.6% and 97.9% for ethambutol, respectively. Agreement between NRA and proportion method were 99.1%, 97.3%, 93.8%, 95.6% for isoniazid, rifampicin, streptomycin and ethambutol, respectively. Conclusion: NRA is easier, inexpensive and reliable method for susceptibility testing of Mycobacterum tuberculosis for isoniazid and rifampicin, the two most important drugs for the treatment of tuberculosis. The reduction in susceptibility testing time, and higher sensitivity and specificity of NRA method is of fundamental importance in detecting MDR-TB. SAARC Journal of Tuberculosis, Lung Diseases & HIV/AIDS; 2012; IX(2) 5-8 DOI: http://dx.doi.org/10.3126/saarctb.v9i2.7971
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Al-Madhagi, Anwar K., Khaled A. Al-Moyed, and Ahmed M. Al-Haddad. "Resistance of anti-tuberculosis drugs among pulmonary tuberculosis patients in Yemen." Journal of the Faculty of Medicine Baghdad 55, no. 3 (July 1, 2013): 250–53. http://dx.doi.org/10.32007/jfacmedbagdad.553625.
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Background: Tuberculosis (TB) is still a major global public health problem worldwide. The global prevalence of Mycobacterium (M tuberculosis) infection has been estimated in 32% of the world population with more than 8 million new cases diagnosed each year.
Materials and Methods: A total of 192 M tuberculosis complex isolates were collected from patients with positive sputum smear who had been treated previously with the four main anti- tuberculosis drugs for more than two months. The isolates were identified by their colonial morphology, pigmentation, shapes on Ziehl-Neelsen smears, growth on Löwenstein-Jenson medium and biochemical tests as niacin and nitrate tests. A proportional method was used for the in vitro drug susceptibility testing.Objective: To estimate drug resistance among previously treated tuberculosis patients, focusing on multi-drug resistant strains at two time intervals (2002 and 2009) in Yemen.
Results: Of the 192 M tuberculosis complex tested isolates, 55 (28.7%) were resistant to one or more drug; 20 (10.4%) were resistant to one drug, 13 (6.8%) to two drugs, 13 (6.8%) to three drugs and 9 (4.7%) to four drugs. Regarding the resistance to an individual drug, out of 192 tested isolates, 36 (18.7%) were resistant to rifampicin, 34 (17.7%) to isoniazid, 33 (17.2%) to ethambutol and 18 (9.4%) to streptomycin and these results were without a statistical significance. The incidence of multidrug resistance against rifampicin and isoniazid with or without other drugs was 13.5% among the tested M tuberculosis complex strains and this result was also without a statistical significance.
Conclusion: The results of this study revealed nearly similar drug resistance patterns for the tested isolates in comparison with previous findings of 2002 and the emergence of more multi-drug resistance M tuberculosis complex strains after a time interval in Yemen.
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Suhas Kulkarni, Gauri, and Chetan Mahajan. "Resistance to Isoniazid and Rifampicin and Factors Associated with Resistance Among MDR TB Patients Visiting DOTS PLUS Site." MVP Journal of Medical Sciences 4, no. 1 (May 22, 2017): 14. http://dx.doi.org/10.18311/mvpjms/0/v0/i0/14782.
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Introduction: Multidrug-Resistant (MDR-TB) is defined as M tuberculosis resistant to isoniazid and rifampicin with or without resistant to other drugs. Drug resistant TB is known to occur from time of introduction of antituberculosis drugs. MDR TB has become a significant health problem and an obstacle to effective TB control1. Resistance of M. tuberculosis to anti- TB drugs is caused by chromosomal mutations in genes encoding drug targets. Multidrug-resistant strains of M. tuberculosis (MDR-TB) evolve due to sequential accumulation of mutations in target genes. The WHO cites TB as the single most important fatal infection, with around 8.8 million new cases and 1.4 millions deaths per year, 95% in developing countries. According to Global TB report 2015 data of MDR TB as follows2. 50% successfully completed treatment (cure or treatment completed), 16% died, 16% defaulters, 10% treatment failure, 8% without outcome. The MDR-TB is also threatening World Health Organization’s target of tuberculosis elimination by 2050. Study conducted by NIRT and NTI suggest MDR level of 1% to 3% in new cases and around 12% in retreatment cases and revealed an overall emergence to rifampicin in only 2% of patients, despite a high level (18%) of initial resistance to isoniazid, either alone in or in combination with other anti tuberculosis drugs+. Aims and Objectives: 1. To Study the drug resistance to isoniazid and rifampicin among the MDR TB patients visiting DOTS PLUS CENTRE. 2. To Study the various factors associated with resistance to Isoniazid and rifampicin among MDR TB patients. Material and Methods: Present study was conducted at DOTS PLUS Centre in tertiary health care centre. Total 140 of newly diagnosed cases of MDR-TB were included in the present study after satisfying the inclusion and exclusion criteria. Written informed consent was taken from the study participants. Patients’demographic details were noted such as name, age, sex, occupation, socioeconomic status, education. Patients were asked detailed history about smoking, alcohol, tobacco chewing, diabetes mellitus, hypertension, COPD (chronic obstructive pulmonary disease), HIV Status. Details about past history of tuberculosis, treatment history were noted, If the patient was found to have defaulted previous antituberculous treatment, detailed evaluation was done to find out reasons for defaulting the treatment, History of MDR TB contact was noted. The drug resistance pattern of isoniazid and rifampicin was noted. Results: Of the 140 drug resistance tuberculosis patients, MDR Pulmonary tuberculosis was more common in economically productive age group of 21-40 years, distribution of male (60%) and female (40%), 35% patients were found to have defaulted previous antituberculous treatment. Main reasons for defaulting were, becoming asymptomatic, feeling better 40.8% followed by medication side effects 32.65% Conclusion: Isoniazid and rifampicin resistance (74.28%)is more than rifampicin monoresistance (25.72%), there is significant association between addiction of patient and defaulting the previous antituberculous treatment, analysis of patients various factors for drug resistance showed that MDR-TB is more commonly seen in males, age between 21 to 40 years, low socioeconomic status, past history of ATT, Alcoholic and tobacco user.
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Solari, L., D. Santos-Lazaro, and Z. M. Puyen. "Mutations in Mycobacterium tuberculosis Isolates with Discordant Results for Drug-Susceptibility Testing in Peru." International Journal of Microbiology 2020 (April 7, 2020): 1–5. http://dx.doi.org/10.1155/2020/8253546.
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Evaluation of resistance to antituberculosis drugs is routinely performed with genotypic or phenotypic methods; however, discordance can be seen between these different methodologies. Our objective was to identify mutations that could explain discordant results in the evaluation of susceptibility to rifampicin and isoniazid between molecular and phenotypic methods, using whole genome sequencing (WGS). Peruvian strains showing sensitive results in the GenoType MTBDRplus v2.0 test and resistant results in the proportions in the agar-plaque test for isoniazid or rifampin were selected. Discordance was confirmed by repeating both tests, and WGS was performed, using the Next Generation Sequencing methodology. Obtained sequences were aligned “through reference” (genomic mapping) using the program BWA with the algorithm “mem”, using as a reference the genome of the M. tuberculosis H37Rv strain. Discordance was confirmed in 14 strains for rifampicin and 21 for isoniazid, with 1 strain in common for both antibiotics, for a total of 34 unique strains. The most frequent mutation in the rpoB gene in the discordant strains for rifampicin was V170F. The most frequent mutations in the discordant strains for isoniazid were katG R463L, kasA G269S, and Rv1592c I322V. Several other mutations are reported. This is the first study in Latin America addressing mutations present in strains with discordant results between genotypic and phenotypic methods to rifampicin and isoniazid. These mutations could be considered as future potential targets for genotypic tests for evaluation of susceptibility to these drugs.
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Caminero, Jose, and Charles Daley. "Management of Multidrug-Resistant Tuberculosis." Seminars in Respiratory and Critical Care Medicine 39, no. 03 (June 2018): 310–24. http://dx.doi.org/10.1055/s-0038-1661383.
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AbstractDrug-resistant strains of Mycobacterium tuberculosis pose a major threat to global tuberculosis control. Despite the availability of curative antituberculosis therapy for nearly half a century, inappropriate and inadequate treatment of tuberculosis, as well as unchecked transmission of M. tuberculosis, has resulted in alarming levels of drug-resistant tuberculosis. The World Health Organization (WHO) estimates that there were 600,000 cases of multidrug-resistant tuberculosis (MDR-TB)/rifampin-resistant (RR) tuberculosis in 2016, defined as strains that are resistant to at least isoniazid and rifampicin. Globally, WHO estimates that 4.1% of new tuberculosis cases and 19% of retreatment cases have MDR-TB. By the end of 2016, 123 countries had reported at least one case of extensively drug-resistant strains, which are MDR-TB strains that have acquired additional resistance to fluoroquinolones and at least one second-line injectable. It is estimated that only 22% of all MDR-TB cases are currently receiving therapy. This article reviews the management of MDR/RR-TB and updates recommendations regarding the use of shorter course regimens and new drugs.
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Al-Mutairi, Noura M., Suhail Ahmad, Eiman Mokaddas, and Sahal Al-Hajoj. "First insights into the phylogenetic diversity of Mycobacterium tuberculosis in Kuwait and evaluation of REBA MTB-MDR assay for rapid detection of MDR-TB." PLOS ONE 17, no. 10 (October 20, 2022): e0276487. http://dx.doi.org/10.1371/journal.pone.0276487.
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Early detection of Mycobacterium tuberculosis (Mtb) in clinical specimens, its susceptibility to anti-TB drugs and disruption of infection transmission to new hosts are essential components for global tuberculosis (TB) control efforts. This study investigated major Mtb genotypes circulating in Kuwait and evaluated the performance of REBA MTB-MDR (REBA) test in comparison to GenoType MTBDRplus (gMTBDR+) assay for rapid detection of resistance of Mtb to isoniazid and rifampicin (MDR-TB). M. tuberculosis isolates (n = 256) originating predominantly from expatriate patients during a 6-month period were tested by spoligotyping and a dendrogram was created by UPGMA using MIRU-VNTRplus software. Phenotypic drug susceptibility testing (DST) was performed by MGIT 960 system. Genotypic DST for isoniazid and rifampicin was done by REBA and gMTBDR+ assays. Spoligotyping assigned 188 (73.4%) isolates to specific spoligotype international type (SIT) while 68 isolates exhibited orphan patterns. All major M. tuberculosis lineages were detected and EAI, CAS and Beijing families were predominant. Phylogenetic tree showed 131 patterns with 105 isolates exhibiting a unique pattern while 151 isolates clustered in 26 patterns. Fifteen isolates were resistant to one/more drugs. REBA and gMTBDR+ detected isoniazid resistance in 11/12 and 10/12 and rifampicin resistance in 4/5 and 4/5 resistant isolates, respectively. The diversity of SIT patterns are highly suggestive of infection of most expatriate patients with unique Mtb strains, likely acquired in their native countries before their arrival in Kuwait. Both, REBA and gMTBDR+ assays performed similarly for detection of resistance of Mtb to isoniazid and rifampicin for rapid detection of MDR-TB.
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MADANIA, AMMAR, MAYA HABOUS, HANA ZARZOUR, IFAD GHOURY, and BAREA HEBBO. "Characterization of Mutations Causing Rifampicin and Isoniazid Resistance of Mycobacterium tuberculosis in Syria." Polish Journal of Microbiology 61, no. 1 (2012): 23–32. http://dx.doi.org/10.33073/pjm-2012-003.
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In order to characterize mutations causing rifampicin and isoniazid resistance of M. tuberculosis in Syria, 69 rifampicin resistant (Rif(r)) and 72 isoniazid resistant (Inh(r)) isolates were screened for point mutations in hot spots of the rpoB, katG and inhA genes by DNA sequencing and real time PCR. Of 69 Rif(r) isolates, 62 (90%) had mutations in the rifampin resistance determining region (RRDR) of the rpoB gene, with codons 531 (61%), 526 (13%), and 516 (8.7%) being the most commonly mutated. We found two new mutations (Asp516Thr and Ser531Gly) described for the first time in the rpoB-RRDR in association with rifampicin resistance. Only one mutation (Ile572Phe) was found outside the rpoB-RRDR. Of 72 Inh(r) strains, 30 (41.6%) had a mutation in katGcodon315 (with Ser315Thr being the predominant alteration), and 23 (32%) harbored the inhA(-15C-->T) mutation. While the general pattern of rpoB-RRDR and katG mutations reflected those found worldwide, the prevalence of the inhA(-15C-->T mutation was above the value found in most other countries, emphasizing the great importance of testing the inhA(-15C-->T) mutation for prediction of isoniazid resistance in Syria. Sensitivity of a rapid test using real time PCR and 3'-Minor groove binder (MGB) probes in detecting Rif(r) and Inh(r) isolates was 90% and 69.4%, respectively. This demonstrates that a small set of MGB-probes can be used in real time PCR in order to detect most mutations causing resistance to rifampicin and isoniazid.
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Salina, T. Yu, and T. I. Morozova. "Prevalence and Patterns of Gene Mutations Associated with <i>M. tuberculosis</i> Resistance to Isoniazid and Rifampicin in Patients with Different Clinical Manifestations of Tuberculosis." Tuberculosis and Lung Diseases 101, no. 1 (March 4, 2023): 28–33. http://dx.doi.org/10.58838/2075-1230-2023-101-1-28-33.
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The objective: to study prevalence and patterns of mutations in the katG, inhA, ahpC, rpoB genes associated with Mycobacterium tuberculosis (MTB) resistance to isoniazid (H) and rifampicin (R) in patients with various clinical manifestations of pulmonary tuberculosis (TB).Subjects and Methods. 441 sputum samples collected in tuberculosis patients were tested using biological microchips. Tests were carried out in Group 1 - patients with confirmed bacterial excretion (n = 256) and in Group 2 (n = 185) - patients without bacterial excretion. The same patients were enrolled in Group 3 - patients with acute progressing tuberculosis (n = 52) and Group 4 (n = 99) - patients with localized tuberculosis.Results. In Group 1, DNA of Mycobacterium tuberculosis was found in 79.3% of patients, in Group 2 - in 57.8%. Among all samples, mutations in the genes encoding resistance to isoniazid were detected in 15.5%, resistance to rifampicin - in 58.1%. Resistance to isoniazid was more often caused by mutations in the katG gene (49%) versus the inhA (29%) and ahpC (4.2%) genes. We found 13 most common types of mutations in the rpoB gene associated with resistance to rifampicin. The dominant mutations in both groups were Seu531->Leu mutations - 19.7% in Group 1 and 24.3% in Group 2. In Group 1, mutations in the katG gene (53.7%) were observed more often than mutations in the inhA gene (27.7%). In Group 3, mutations in the katG gene were registered in 30.8%, in the inhA gene - in 25%. There were no statistically significant differences in patterns of mutations in the katG, inhA, ahpC, rpoB genes between Groups 1, 2 and 3, 4. Thus, patients without bacterial excretion and patients with localized tuberculosis are a hidden dangerous reservoir of tuberculous mycobacteria with multiple drug resistance to rifampicin and drug resistance to isoniazid.
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Krajewska-Wędzina, Monika, Anna Zabost, Ewa Augustynowicz-Kopeć, Marcin Weiner, and Krzysztof Szulowski. "Evaluation of susceptibility to antimycobacterial drugs in Mycobacterium tuberculosis complex strains isolated from cattle in Poland." Journal of Veterinary Research 61, no. 1 (March 1, 2017): 23–26. http://dx.doi.org/10.1515/jvetres-2017-0003.
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Abstract Introduction: Tuberculosis is a highly infectious disease affecting humans and animals. It is caused by the Mycobacterium tuberculosis complex (MTBC) – Mycobacterium bovis and Mycobacterium caprae, which are aetiological factors of bovine tuberculosis (bTB). In Poland, the bTB eradication programme exists. Animals diagnosed with tuberculosis are in the majority of cases not treated, but removed from their herd and then sanitary slaughtered. Material and Methods: In total, 134 MTBC strains isolated from cattle in Poland were subjected to microbiological analysis. The resistance phenotype was tested for first-line antimycobacterial drugs used in tuberculosis treatment in humans: streptomycin, isoniazid, rifampicin, ethambutol, and pyrazinamide. The strains were isolated from tissues collected post mortem, so the test for drug resistance fulfilled only epidemiological criterion. Results: The analysis of drug-resistance of MTBC strains revealed that strains classified as M. bovis were susceptible to 4 antimycobacterial drugs: isoniazid, rifampicin, streptomycin, and ethambutol, and resistant to pyrazynamide. The strains classified as M. caprae were sensitive to all tested drugs. Conclusion: The results indicate that despite enormously dynamic changes in mycobacterial phenotype, Polish strains of MTBC isolated from cattle have not acquired environmental resistance. The strains classified as M. bovis are characterised by natural resistance to pyrazinamide, which is typical for this species.
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Admassu, Wasihun, Birhanu Ayelign, Gemeda Abebe, and Mulualem Tadesse. "Detection of Mycobacterium tuberculosis and rifampicin resistance by Xpert® MTB/RIF assay among presumptive tuberculosis cases at Jimma University Medical Center, Southwest Ethiopia." PLOS ONE 17, no. 1 (January 27, 2022): e0262929. http://dx.doi.org/10.1371/journal.pone.0262929.
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Background Rapid diagnosis of tuberculosis (TB) and detection of drug resistance are very important for timely and appropriate management of patients. Xpert MTB/RIF assay is approved for use in TB and rifampicin-resistance diagnosis. However, data are limited on the impact of Xpert MTB/RIF assay under routine clinical settings with a heterogeneous group of patients and sample types in Ethiopia. Methods A retrospective study was carried out in 2220 presumptive TB cases at Jimma University Medical Center. Data were gathered from the registration logbook using formatted data extraction tools and double entered to epidata version 3.1 and further transported to SPSS version 20 for analysis. Associations were determined using the Chi-square test and P-value <0.05 was considered statistically significant. Results Of 2220 cases enrolled, 1665 (75%) were adults and the remaining 555 (25%) were children aged less than 14 years. The majority, 1964 (88.46%), had pulmonary manifestation and 256 (11.54%) had extrapulmonary involvements. The overall, frequency of Mycobacterium tuberculosis (MTB) was 9.3% (206/2220), among this 10.27% (171/1665) and 6.3% (35/555) were adults and children, respectively. M. tuberculosis was detected from 171 (8.75%) of pulmonary patients and 35 (13.28%) of extrapulmonary manifested patients. Out of 206 M. tuberculosis positive cases, 7(3.4%) were rifampicin-resistant: four from pulmonary tuberculosis (PTB) patients and three from EPTB patients. In the Chi-square test, the age group of 15–24 years, previous history of TB, pus/lymph node sample, and being HIV positive were significantly associated with TB positivity by Xpert MTB/RIF (P-value <0.001). Conclusion These data suggest that the overall frequency of M. tuberculosis and rifampicin resistance was found to be relatively low compared to the previous reports in Ethiopia. Nevertheless, better diagnostic tools and approaches are still vital to halt the burden of TB and drug-resistant TB in the country.
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Kalani, Komal, Vinita Chaturvedi, Priyanka Trivedi, Sudeep Tondon, and Santosh Kumar Srivastava. "Dihydroartemisinin and its Analogs: A New Class of Antitubercular Agents." Current Topics in Medicinal Chemistry 19, no. 8 (June 3, 2019): 594–99. http://dx.doi.org/10.2174/1568026619666190304142802.
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Background: Tuberculosis is one of the leading causes of mortality worldwide. Resistance against the frontline anti-tubercular drugs has worsened the already alarming situation, which requires intensive drug discovery to develop new, more effective, affordable and accessible anti-tubercular agents possessing novel modes of action. Objective: Chemical transformation of dihydroartemisinin for anti-tubercular lead optimization. Methods: Dihydroartemisinin, a metabolite of artemisinin was chemically converted into eight acyl derivatives and were evaluated for anti-tubercular potential against H37Rv virulent strain of Mycobacterium tuberculosis by agar-based proportion assay. Further, synergistic activity of 12-O-m-anisoyl dihydroartemisinin was also studied with the front-line anti-TB drugs, isoniazid and rifampicin. Results: The results showed that all the derivatives were active but out of eight, 12-O-m-anisoyl dihydroartemisinin and 12-O-p-anisoyl dihydroartemisinin were significantly active (MIC 25.0 µg/mL). In synergistic activity evaluation, the 12-O-m-anisoyl dihydroartemisinin derivative showed reduction in MIC (by 1/8th, i.e. 3.12 µg/mL and that of rifampicin by ¼th, i.e. 0.05 µg/mL) with the front-line anti-TB drug, rifampicin. The sumfractional inhibitory concentration (Σ FIC) was 0.375. Conclusion: These results suggested a synergistic effect of the 12-O-m-anisoyl dihydroartemisinin with rifampicin and established its base for the development of anti-tubercular agents from an in-expensive and non-toxic natural product. To the best of our knowledge this is the first ever report on the anti-tubercular potential of dihydroartemisinin and its derivatives.
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Umpeleva, T. V., E. A. Mazurina, D. V. Vakhrusheva, and N. I. Eremeeva. "Comparison of Different Methods for Drug Susceptibility Testing of Mycobacterium tuberculosis to Rifampicin." Tuberculosis and Lung Diseases 100, no. 1 (February 11, 2022): 41–48. http://dx.doi.org/10.21292/2075-1230-2022-100-1-41-48.
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The objective: to compare results of drug susceptibility testing to rifampicin by molecular genetic methods and phenotypic tests of Mycobacterium tuberculosis isolates obtained from clinical specimens of tuberculosis patients.Subjects and Methods. 915 samples of M. tuberculosis DNA and 426 cultures were used in this study. Genotypic tests (TB-TEST (BIOCIP-IMB, Russia), GenoType MTBDRplusV2) and phenotypic technologies (absolute concentration method, Bactec MGIT 960 system, Sensititre Myco TB kit) were used.Results. A high percentage (98.7%; CI 97.7-99.7%) of confirmation of the results of the molecular genetic test (TB-TEST) by the phenotypic test (absolute concentration method) was demonstrated. In some cases, the Bactec MGIT 960 system as well as the absolute concentration method were shown to produce false negative results of rifampicin resistance in some cases.
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Diriba, Getu, Abebaw Kebede, Habteyes Hailu Tola, Ayinalem Alemu, Bazezew Yenew, Shewki Moga, Desalegn Addise, et al. "Utility of line probe assay in detecting drug resistance and the associated mutations in patients with extrapulmonary tuberculosis in Addis Ababa, Ethiopia." SAGE Open Medicine 10 (January 2022): 205031212210982. http://dx.doi.org/10.1177/20503121221098241.
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Introduction: Molecular tests allow rapid detection of Mycobacterium tuberculosis and drug resistance in a few days. Identifying the mutations in genes associated with drug resistance may contribute to the development of appropriate interventions to improve tuberculosis control. So far, there is little information in Ethiopia about the diagnostic performance of line probe assay (LPA) and the M. tuberculosis common gene mutations associated with drug resistance in extrapulmonary tuberculosis. Thus, this study aimed to assess the frequency of drug resistance-associated mutations in patients with extrapulmonary tuberculosis (EPTB) and to compare the agreement and determine the utility of the genotypic in the detection of drug resistance in Addis Ababa, Ethiopia. Methods: A cross-sectional study was conducted on stored M. tuberculosis isolates. The genotypic and phenotypic drug susceptibility tests were performed using LPA and BACTEC-MGIT-960, respectively. The common mutations were noted, and the agreement and the utility of the LPA were determined using the BACTEC-MGIT-960 as a gold standard. Results: Of the 151 isolates, the sensitivity and specificity of MTBDR plus in detecting isoniazid resistance were 90.9% and 100%, respectively. While for rifampicin, it was 100% and 99.3% for sensitivity and specificity, respectively. The katG S315Tl was the most common mutation observed in 85.7% of the isoniazid-resistant isolates. In the case of rifampicin, the most common mutation (61.9%) was observed at position rpoB S531L. Mutations in the gyrA promoter region were strongly associated with Levofloxacin and Moxifloxacin resistance. Conclusion: Line probe assay has high test performance in detecting resistance to anti-TB drugs in EPTB isolates. The MTBDR plus test was slightly less sensitive for the detection of isoniazid resistance as compared to the detection of rifampicin. The most prevalent mutations associated with isoniazid and rifampicin resistance were observed at katG S315Tl and rpoB S531L respectively. Besides, all the fluoroquinolone-resistant cases were associated with gyrA gene. Finally, a validation study with DNA sequencing is recommended.
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Kazakova, Oxana, Roxana Racoviceanu, Anastasiya Petrova, Marius Mioc, Adrian Militaru, Lucreția Udrescu, Mihai Udrescu, et al. "New Investigations with Lupane Type A-Ring Azepane Triterpenoids for Antimycobacterial Drug Candidate Design." International Journal of Molecular Sciences 22, no. 22 (November 21, 2021): 12542. http://dx.doi.org/10.3390/ijms222212542.
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Twenty lupane type A-ring azepano-triterpenoids were synthesized from betulin and its related derivatives and their antitubercular activity against Mycobacterium tuberculosis, mono-resistant MTB strains, and nontuberculous strains Mycobacterium abscessus and Mycobacterium avium were investigated in the framework of AToMIc (Anti-mycobacterial Target or Mechanism Identification Contract) realized by the Division of Microbiology and Infectious Diseases, NIAID, National Institute of Health. Of all the tested triterpenoids, 17 compounds showed antitubercular activity and 6 compounds were highly active on the H37Rv wild strain (with MIC 0.5 µM for compound 7), out of which 4 derivatives also emerged as highly active compounds on the three mono-resistant MTB strains. Molecular docking corroborated with a machine learning drug-drug similarity algorithm revealed that azepano-triterpenoids have a rifampicin-like antitubercular activity, with compound 7 scoring the highest as a potential M. tuberculosis RNAP potential inhibitor. FIC testing demonstrated an additive effect of compound 7 when combined with rifampin, isoniazid and ethambutol. Most compounds were highly active against M. avium with compound 14 recording the same MIC value as the control rifampicin (0.0625 µM). The antitubercular ex vivo effectiveness of the tested compounds on THP-1 infected macrophages is correlated with their increased cell permeability. The tested triterpenoids also exhibit low cytotoxicity and do not induce antibacterial resistance in MTB strains.
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Sanzhakov, M. A., O. M. Ipatova, V. N. Prozorovskiy, N. V. Medvedeva, and T. I. Torkhovskaya. "Interaction of rifampicin embedded in phospholipid nanoparticles with blood plasma lipoproteins." Biomeditsinskaya Khimiya 60, no. 3 (2014): 348–53. http://dx.doi.org/10.18097/pbmc20146003348.
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The drug formulations of antituberculous remedy rifampicin in nanoparticles less than 30 nm based on soy phosphatidylcholine and sodium oleate was elaborated in Institute of Biomedical Chemistry. The distribution of rifampicin in blood plasma fractions after incubation with this formulation and with free rifampicin was studied. This goal was stimulated by the literature data about activation of macrophages LDL receptors in cases of M. tuberculosis infection. Plasma was incubated 30 min with free rifampicin or rifampicin encapsulated into the nanoformulation followed by ultracentrifugation and subsequent rifampicin determination by HPLC in lipoprotein fractions. In the case of free rifampicin it appeared mainly in the plasma protein fraction and in HDL (41% and 38%, correspondentely). But after incubation of rifampicin in nanoparticles the drug redistribution was observed. Its proportion in these factions decreased 2-3-fold, and it was found mainly in LDL (60% as compared with 21% for free rifampicin). The increased association of rifampicin encapsulated into phospholipid nanoparticles with LDL is considered as facilitating factor for macrophages delivery and thus for antituberculosis efficiency as well
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Mohamad, Suriyati, Nur Najihah Ismail, Hasnah Osman, Habibah A Wahab, and Thaigarajan Parumasivam. "In Vitro Interactions of Costus speciosus (J. Koenig) Sm., Cymbopogon citratus (Dc. Ex Nees) Stapf. and Tabernaemontana coronaria (L.) Willd. with First-Line Anti-Tuberculosis Drugs Against Mycobacterium tuberculosis H37rv." Malaysian Journal of Pharmaceutical Sciences 19, no. 2 (November 23, 2021): 171–78. http://dx.doi.org/10.21315/mjps2021.19.2.11.
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Global tuberculosis (TB) burden underscores the importance of developing new effective anti-TB drugs. This study was concerned with prospecting for potential anti-TB agents from Malaysian medicinal plants. In our previous study, we have reported that n-hexane fractions of Costus speciosus (C. speciosus) (J. Koening) Sm., Cymbopogon citratus (C. citratus ) (DC.) Stapf. and Tabernaemontana coronaria (T. coronaria) (Jacq.) posses promising anti-TB activity against Mycobacterium tuberculosis (M. tuberculosis) H37Rv with minimum inhibitory concentrations (MICs) of 200–100 µg/mL. This study aimed to investigate the interactions of these active fractions with first-line anti-TB drugs (isoniazid, rifampicin, ethambutol and streptomycin) against M. tuberculosis H37Rv using the microdilution checkerboard method. C. citratus (stem-rhizome) n-hexane fraction exhibited synergism with all drugs except ethambutol which showed additive interaction. Synergistic was also observed when C. speciosus (stem-flower) n-hexane and T. coronaria (leaf) n-hexane fractions in combination with rifampicin. C. speciosus (stem-flower) n-hexane and T. coronaria (leaf) n-hexane exhibited additive interaction with isoniazid, ethambutol and streptomycin. Hence, these active plants are worthy of further investigations for the discovery of anti-TB drug leads.
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Sarkar, Sampa, and Dhiman Sarkar. "Potential Use of Nitrate Reductase as a Biomarker for the Identification of Active and Dormant Inhibitors of Mycobacterium tuberculosis in a THP1 Infection Model." Journal of Biomolecular Screening 17, no. 7 (May 9, 2012): 966–73. http://dx.doi.org/10.1177/1087057112445485.
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The development of a macrophage-based, antitubercular high-throughput screening system could expedite discovery programs for identifying novel inhibitors. In this study, the kinetics of nitrate reduction (NR) by Mycobacterium tuberculosis during growth in Thp1 macrophages was found to be almost parallel to viable bacilli count. NR in the culture medium containing 50 mM of nitrate was found to be optimum on the fifth day after infection with M. tuberculosis. The signal-to-noise (S/N) ratio and Z-factor obtained from this macrophage-based assay were 5.4 and 0.965, respectively, which confirms the robustness of the assay protocol. The protocol was further validated by using standard antitubercular inhibitors such as rifampicin, isoniazid, streptomycin, ethambutol, and pyrazinamide, added at their IC90 value, on the day of infection. These inhibitors were not able to kill the bacilli when added to the culture on the fifth day after infection. Interestingly, pentachlorophenol and rifampicin killed the bacilli immediately after addition on the fifth day of infection. Altogether, this assay protocol using M. tuberculosis–infected Thp-1 macrophages provides a novel, cost-efficient, robust, and easy-to-perform screening platform for the identification of both active and hypoxic stage-specific inhibitors against tuberculosis.
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Kumar, Vipul, Jyoti Yadav, Aparna Parmar, Ritu Aggarwal, and K. B. Gupta. "Study of rifampicin resistance among newly diagnosed pulmonary tuberculosis patients with type 2 diabetes mellitus: a prospective observational study." International Journal of Research in Medical Sciences 9, no. 7 (June 25, 2021): 2035. http://dx.doi.org/10.18203/2320-6012.ijrms20212526.
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Background: Rifampicin (RIF) resistance in new cases of pulmonary tuberculosis is a matter of concern. Diabetes Mellitus triples the risk of developing tuberculosis. Early detection of TB and its resistance status in diabetics can help in improving the care and treatment outcomes of both diseases.Methods: It was a prospective study conducted from February 2019 to March 2020 in PGIMS, Rohtak on 50 patients of DM with newly diagnosed Pulmonary TB. Rifampicin resistance was detected by CBNAAT on sputum, induced sputum and BAL samples. Results: Mean age of study subjects was 51.24±10.421 (in years) with M: F ratio of 3:1 and maximum patients in 50-59 age group. The average BMI in patients was 22.49±2.42 kg/m2. The most common presenting complaint was cough (92%) followed by fever (68%). Mean serum HBA1c was 9.66±2.24 and mean FBS and PPBS was 195.81±59.08 and 302.02±99.01 mg/dl respectively. Out of 36 cases who gave sputum, 29 (80.55%) were detected rifampicin sensitive and 7 (19.44%) were rifampicin resistant whereas out of 11 who were induced, 9 (81.8%) were rifampicin sensitive and 2(18.18%) were rifampicin resistant. Out of 3 cases detected by BAL CBNAAT, 2 (66.6%) were rifampicin sensitive and 1 (33.33%) was rifampicin resistant. Overall, 10(20%) patients were detected Rifampicin resistant by CBNAAT.Conclusions: We found that TB-DM patients had a higher proportion of drug resistance (20%), so DM should be considered as an independent risk factor for MDR-TB and effective measures are required for early diagnosis of MDR-TB.
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Dey, Abhinav, Amit Kumar Verma, and Dipankar Chatterji. "Role of an RNA polymerase interacting protein, MsRbpA, from Mycobacterium smegmatis in phenotypic tolerance to rifampicin." Microbiology 156, no. 3 (March 1, 2010): 873–83. http://dx.doi.org/10.1099/mic.0.033670-0.
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Rifampicin and its derivatives are at the forefront of the current standard chemotherapeutic regimen for active tuberculosis; they act by inhibiting the transcription activity of prokaryotic RNA polymerase. Rifampicin is believed to interact with the β subunit of RNA polymerase. However, it has been observed that protein–protein interactions with RNA polymerase core enzyme lead to its reduced susceptibility to rifampicin. This mechanism became more diversified with the discovery of RbpA, a novel RNA polymerase-binding protein, in Streptomyces coelicolor that could mitigate the effect of rifampicin on RNA polymerase activity. MsRbpA is a homologue of RbpA in Mycobacterium smegmatis. On deciphering the role of MsRbpA in M. smegmatis we found that it interacts with RNA polymerase and increases the rifampicin tolerance levels, both in vitro and in vivo. It interacts with the β subunit of RNA polymerase. However, it was found to be incapable of rescuing rifampicin-resistant RNA polymerases in the presence of rifampicin at the respective IC50.
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Do, Thi Thuy, Jerónimo Rodríguez-Beltran, Esmeralda Cebrián-Sastre, Alexandro Rodríguez-Rojas, Alfredo Castañeda-García, and Jesús Blázquez. "Inactivation of a New Potassium Channel Increases Rifampicin Resistance and Induces Collateral Sensitivity to Hydrophilic Antibiotics in Mycobacterium smegmatis." Antibiotics 11, no. 4 (April 12, 2022): 509. http://dx.doi.org/10.3390/antibiotics11040509.
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Rifampicin is a critical first-line antibiotic for treating mycobacterial infections such as tuberculosis, one of the most serious infectious diseases worldwide. Rifampicin resistance in mycobacteria is mainly caused by mutations in the rpoB gene; however, some rifampicin-resistant strains showed no rpoB mutations. Therefore, alternative mechanisms must explain this resistance in mycobacteria. In this work, a library of 11,000 Mycobacterium smegmatis mc2 155 insertion mutants was explored to search and characterize new rifampicin-resistance determinants. A transposon insertion in the MSMEG_1945 gene modified the growth rate, pH homeostasis and membrane potential in M. smegmatis, producing rifampicin resistance and collateral susceptibility to other antitubercular drugs such as isoniazid, ethionamide and aminoglycosides. Our data suggest that the M. smegmatis MSMEG_1945 protein is an ion channel, dubbed MchK, essential for maintaining the cellular ionic balance and membrane potential, modulating susceptibility to antimycobacterial agents. The functions of this new gene point once again to potassium homeostasis impairment as a proxy to resistance to rifampicin. This study increases the known repertoire of mycobacterial ion channels involved in drug susceptibility/resistance to antimycobacterial drugs and suggests novel intervention opportunities, highlighting ion channels as druggable pathways.
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Bondarenko, V. N., V. A. Shtanze, and L. V. Zolotukhina. "CHARACTERISTICS OF <i>M.TUBERCULOSIS</i> DRUG RESISTANCE DETERMINED BY MOLECULAR GENETIC AND PHENOTYPIC METHODS." Health and Ecology Issues, no. 3 (September 28, 2018): 61–66. http://dx.doi.org/10.51523/2708-6011.2018-15-3-12.
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Objective: to determine the genetic and phenotypic drug resistance of M . tuberculosis to first-line and second-line anti-TB drugs. Material and methods. Gene mutations in 247 strains of M. tuberculosis (MBT) associated with drug resistance to isoniazid, rifampicin, fluoroquinolones, and aminoglycosides were studied. Genetic resistance of a tuberculosis causative agent was determined by means of LPA (GenoType® MTBDRsl MTBDRplus and MTBDRsl, ver.2.0). The results of the study are confirmed by the determination of phenotypic drug resistance in the automated system BACTEC ™ MGIT ™ 960. Results. The drug resistant MBT strains circulating around Gomel region have been determined, and the high reliability of molecular and genetic determination of MBT drug resistance has been confirmed by microbiological methods (isoniazid and rifampicin - 97.2 %, fluoroquinolones - 85.1 %, aminoglycosides - 92.3 %). A considerable number of drug resistant MTB strains with gene mutations (45.1 %) which are not included in the GenoType® MTBDRsl system were detected. Conclusion. Тhe considerable genetic variability of drug-resistant MBT strains requires complex application of all the methods of drug resistance testing.
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Darma, Surya, Angga Ambara, Abu Tholib Aman, Luthvia Annisa, Nurrokhman, Titik Nuryastuti, and Tri Wibawa. "High frequency of azole resistant Candida spp. colonization among presumptive multidrug resistant tuberculosis (MDR-TB) patients." PLOS ONE 15, no. 11 (November 19, 2020): e0242542. http://dx.doi.org/10.1371/journal.pone.0242542.
Full textAbstract:
Background Tuberculosis is one of the major causes of death globally. The problems become even more complicated with the rise in prevalence of multidrug resistant tuberculosis (MDR-TB). Many diseases have been reported to occur with tuberculosis making it more difficult to manage. Candida spp., which are yeast-like fungi and a constituent of normal flora in humans, are notoriously reported to be one of the most common opportunistic nosocomial infections. This study aimed to measure the proportion of presumptive MDR-TB patients colonized with Candida spp. and to characterize its susceptibility against azole group antifungal agents. Methods Sputum from presumptive MDR-TB patients were collected and examined for the presence of Mycobacterium tuberculosis and its rifampicin resistant status using GeneXpert. It was further cultured on Sabouroud’s Dextrose Agar (SDA) to isolate the Candida spp. The Candida species were determined using HiCrome™ Candidal Differential Agar. Antifungal susceptibility was tested using microbroth dilution methods. Checkerboard microdilution assays were performed to measure the interaction between rifampicin and fluconazole to C. albicans. Results There were 355 presumptive MDR-TB patients enrolled. A total of 101 (28.4%) patients were confirmed to have M. tuberculosis. There were 113 (31.8%) sputum positive for Candida spp., which corresponded to 149 Candida spp. isolates. Candida albicans was the most frequent (53.7%) species isolated from all patients. The susceptibility of Candida spp. against fluconazole, itraconazole, and ketoconazole were 38.3%, 1.3%, and 10.7% respectively. There was significant association between rifampicin exposure history and susceptibility of Candida albicans against fluconazole (Odds Ratio: 9.96; 95% CI: 1.83–54.19; p <0.01), but not for ketoconazole and itraconazole. The checkerboard microdilution assays showed that rifampicin decreased the fungicidal activity of fluconazole to C. albicans in a dose-dependent manner. Conclusion There was high frequency of azole resistant Candida spp. isolates colonizing the respiratory tract of presumptive MDR-TB patients. This presence might indicate the association of chronic exposure to rifampicin, the main drug for tuberculosis therapy, with the induction of azole resistance.
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Bourama Kané, Korotoumou Wélé Diallo, Boubacar Mami Touré, Oumou Koné, Guédiouma Dembélé, Sow Djénéba Sylla, Modibo Mariko, et al. "Bacilliferous pulmonary tuberculosis of the infant: Report of 3 cases in the pediatrics department of the hospital of Mali." International Journal of Science and Research Archive 1, no. 1 (November 30, 2020): 018–25. http://dx.doi.org/10.30574/ijsra.2020.1.1.0018.
Full textAbstract:
Introduction: Tuberculosis is one of the top ten killers worldwide. In 2015, an estimated one million children developed the disease and 170,000 died from it. We report three cases of pulmonary tuberculosis in infants diagnosed and treated in the pediatric ward of the Mali Hospital. Clinical cases: Observation 1: He is a 7 month old infant hospitalized for fever and weight loss. He received the BCG. There was no notion of TB contagion. On admission, he had poor nutritional status with a Zscore <- 3 and pallor. He had bronchial groans. Xpert / RIF returned positive to M. tuberculosis sensitive to rifampicin. An anti-tuberculosis treatment (2RHZ / 4RH) associated with the transfusion of the globular concentrate at a rate of 20 ml / Kg / 1d over 1 hour and nutritional management have been instituted. After 2 months of treatment, we observed clinical radiological improvement. Xpert control of gastric fluid returned negative. Observation 2: He was an 8-month-old infant hospitalized for fever and weight loss. He did not receive BCG. There was no notion of family storytelling. On admission, he had a poor nutritional status with a Z score <-3. The respiratory rate was 32 cycles / min. There were crackling groans. Direct gastric fluid examination and Xpert / RIF were positive for M. tuberculosis sensitive to rifampicin. He could not be treated because the family requested discharge against all medical advice. Observation 3: He was a 10 month old infant admitted for cough, fever and weight loss. He received the BCG, there was the notion of family contagion. At the entrance, he had a poor nutritional status with a zscore <- 3. He had a polypnea at 45 / min and crackling groans. Direct examination and culture of gastric fluid were positive for rifampicin-sensitive M. tuberculosis. A treatment including oxygen, anti-tuberculosis drugs (2RHZ / 4RH) and nutritional management was initiated. Within 2 months of treatment, we observed clinical and radiological improvement. Direct examination and culture of gastric fluid returned negative. Conclusion: tuberculosis in infants is poorly documented because of unspecific symptoms and difficulties in obtaining bacteriological confirmation. It should be systematically sought in all malnourished infants in endemic countries.
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Khan, Javaid, Najmul Islam, Nadya Ajanee, and Wasim Jafri. "Drug Resistance of Mycobacterium Tuberculosis in Karachi, Pakistan." Tropical Doctor 23, no. 1 (January 1993): 13–14. http://dx.doi.org/10.1177/004947559302300107.
Full textAbstract:
We determined the primary and secondary resistance of isolates of M. tuberculosis to the standard anti-tuberculous drugs in Karachi (Pakistan). Primary resistance to one or more anti-tuberculous drugs was found in 17% of 123 isolates of M. tuberculosis (obtained from patients with no history of previous treatment for tuberculosis). Secondary resistance was found in 36% of 33 isolates (obtained from individuals who had received anti-tuberculous treatment in the past). The drug to which organisms were most commonly resistant was isoniazid (11% primary resistance, 30% secondary resistance). Fifteen per cent of isolates obtained from previously-studied patients showed secondary resistance to rifampicin. We discuss the importance of these findings for tuberculosis treatment and control.