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Kartika Sari, Sri, and Aryati Aryati. "DIAGNOSIS JANGKITAN (INFEKSI) VIRUS DENGUE DENGAN UJI CEPAT (RAPID TEST) IgA ANTI-DENGUE." INDONESIAN JOURNAL OF CLINICAL PATHOLOGY AND MEDICAL LABORATORY 17, no. 2 (March 17, 2018): 81. http://dx.doi.org/10.24293/ijcpml.v17i2.1020.
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Dengue IgM, IgG Capture ELISAs and NS1 Ag ELISA become the most widely used serological methods for dengue diagnosis untilnow. Previous studies reported a possible use of IgA antibodies for dengue virus as a new serologic marker to make dengue infectionactive. In the present study, the performance of IgA anti-dengue rapid test as a new marker of dengue infection was assessed. In thisstudy, the sera were obtained from 30 dengue virus infection patients and 30 non dengue virus infection patients. Thirty dengue pairedsera were collected twice, at the time of hospital admission (acute) and at discharge (convalescent). All sera samples were characterizedusing dengue reference ELISAs (NS1 Ag, Dengue IgM and IgG capture ELISAs). The results of IgA anti-dengue rapid test were comparedwith the corresponding dengue reference tests. The sensitivity and specificity of IgA anti-dengue rapid test respectively were 78.3% (95%CI: 65.5–87.5%), and 73.3% (95% CI: 55,6–85,8%). Meanwhile, from acute sera, sensitivity of IgA anti-dengue rapid test was 83.3%(95% CI: 64.5–93.7), higher than IgM (73.3%, 95% CI: 53.8–87.0), IgG (66.7%, 95% CI: 47.1–82.1) and NS1 Ag ELISAs (60%,95% CI: 40.7–76.8). Positive IgA anti-dengue rapid test results in acute sera was higher in the secondary (91%) than primary infection(57%). IgA anti-dengue rapid test can be considered as a new marker for dengue infection, because it gives a high sensitivity, especiallyin the acute phase and in the secondary infections as well.
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Wu, Shuenn-Jue L., Helene Paxton, Barbara Hanson, Cheryl G. Kung, Timothy B. Chen, Cindy Rossi, David W. Vaughn, Gerald S. Murphy, and Curtis G. Hayes. "Comparison of Two Rapid Diagnostic Assays for Detection of Immunoglobulin M Antibodies to Dengue Virus." Clinical Diagnostic Laboratory Immunology 7, no. 1 (January 1, 2000): 106–10. http://dx.doi.org/10.1128/cdli.7.1.106-110.2000.
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ABSTRACT Two easy-to-use commercial diagnostic assays, a dipstick enzyme-linked immunosorbent assay (ELISA) (Integrated Diagnostics, Baltimore, Md.) and an immunochromatographic card assay (PanBio, Brisbane, Australia) were evaluated for detection of immunoglobulin M (IgM) antibody to dengue virus with an in-house IgM antibody capture microplate ELISA as a reference assay. The dipstick ELISA was based on the indirect-ELISA format using dengue 2 virus as the only antigen and enzyme-labeled goat anti-human IgM antibody as the detector. The total assay time was 75 min. The immunochromatographic card assay was based on the antibody capture format and separately measured both anti-dengue virus IgM and IgG in the same test. Colloidal-gold-labeled anti-dengue virus monoclonal antibody bound with dengue virus 1 to 4 antigen cocktail was the detector, and anti-human IgM and IgG were the capture antibodies. The total assay time was <10 min. Sera from 164 individuals classified as either anti-dengue virus IgM positive (94) or anti-dengue virus IgM negative (70) in the reference microplate ELISA with a dengue virus 1 to 4 antigen cocktail were tested in the two commercial assays. The dipstick ELISA missed 7 of 94 positive samples, for a sensitivity of 92.6%, while the immunochromatographic card assay missed two positive samples, for a sensitivity of 97.9%. Of the 70 negative samples, four were false positive by the dipstick ELISA and two were false positive in the immunochromatographic card assay, resulting in specificities of 94.3 and 97.1%, respectively. Both commercial assays provide sensitive and specific detection of anti-dengue virus IgM antibody and could prove useful in settings where the microplate ELISA is impractical.
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Aryati, Aryati, Puspa Wardhani, Ade Rochaeni, Jeine Stela Akualing, and Usman Hadi. "ANTI DENGUE IGG/IGM RATIO FOR SECONDARY ADULT DENGUE INFECTION IN SURABAYA." INDONESIAN JOURNAL OF CLINICAL PATHOLOGY AND MEDICAL LABORATORY 24, no. 1 (March 29, 2018): 81. http://dx.doi.org/10.24293/ijcpml.v24i1.1161.
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Infeksi Virus Dengue (IVD) dibedakan menjadi infeksi primer dan sekunder berdasarkan respons antibodi yang dihasilkan. Infeksisekunder perlu dibedakan dari infeksi primer karena umumnya menimbulkan manifestasi klinis yang berat. Uji hemaglutinasi inhibisisebagai baku emas untuk menentukan infeksi primer atau sekunder dirasa tidak praktis karena membutuhkan sepasang sera denganselang waktu waktu yang cukup lama. Penelitian ini bertujuan mengetahui cut-off rasio IgG/IgM anti dengue untuk infeksi denguesekunder dewasa di Surabaya. Subjek adalah pasien IVD dengan hasil NS1 dan/atau PCR dengue positif. Rasio IgG/IgM anti-denguediperoleh dari pembagian nilai indeks IgG dan IgM metode ELISA. Nilai cut-off rasio ditentukan berdasarkan kurva ROC. Berdasarkanpola reaktivitas IgM dan IgG ELISA, 19 (31,1%) pasien dikelompokkan sebagai infeksi primer dan 42 (68,9%) infeksi sekunder. HasilPCR didominasi DEN-3. Nilai cut-off optimal rasio IgG/IgM ≥0,927 sebagai peramal infeksi sekunder memiliki kepekaan 66,7% dankekhasan 63,2%. Dianalisis pula nilai cut-off optimal IgM dan IgG anti dengue, yaitu IgM ≥1,515 dan IgG ≥2,034 sebagai peramalinfeksi sekunder memiliki kepekaan dan kekhasans masing-masing 85,7% dan 84,2%; 100% dan 100%. Disimpulkan bahwa rasioIgG/IgM ≥0,927 tidak dapat digunakan sebagai tolok ukur tunggal peramal infeksi sekunder sedangkan cut-off IgG ≥2,034 dapatdipertimbangkan sebagai peramal infeksi sekunder.
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Resna, Resna, Aryati Aryati, Puspa Wardhani, and Erwin Triyono. "NILAI DIAGNOSTIK ANTI DENGUE IgA DAN NS1, SERTA IgM/IgG DI INFEKSI VIRUS DENGUE (The Diagnostic Value of Anti Dengue IgA and Anti Dengue IgM/IgG in Dengue Virus Infection)." INDONESIAN JOURNAL OF CLINICAL PATHOLOGY AND MEDICAL LABORATORY 21, no. 1 (April 15, 2018): 82. http://dx.doi.org/10.24293/ijcpml.v21i1.1264.
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The clinical manifestations of dengue virus infection are varied and thus a specific diagnostic examination is required. Usually antidengueIgM is often used, but the presence in the circulation is 3−8 months long. NS1 is sensitive in the detection of primary infection,whereas IgG is more better used in secondary infection. The examination of anti-dengue IgA as a new marker is estimated to be ableto detect the acute primary and secondary infection, however the diagnostic value of anti-dengue IgA is not much well known for theIndonesian population. This study was done at the Tropical Infectious Disease Ward of Dr. Soetomo Hospital, Surabaya during February– April 2013. The samples consisted of 37 sera from patients infected by dengue virus and 37 sera from those non one (dengue virusinfection patients). The NS1 serum, anti-dengue IgM and anti dengue IgG were examined by ELISA and anti-dengue IgA was examined byan indirect immunochromatography method using Assure@ Dengue IgA Rapid Test (MP Biomedicals Asia Pacific Pte Ltd). The diagnosticvalue was analyzed by 2x2 table with a confidence interval of (CI) 95%. The used gold standards were from the 1997th WHO criteriaand one of the positive dengue serological tests by ELISA (NS1/anti dengue IgM/anti dengue IgG). AUC and anti-dengue IgA cut-off weredetermined by ROC curve. The Diagnostic value of anti-dengue IgA showed a sensitivity and specificity of 83.8% (67.3 to 93.2) and 81.1%(64.3 to 91.4). A positive predictive value of 81.6% (65.1 to 91.7) and a negative predictive value of 83.3% (66.5 to 93.0) was found. Thepositive likelihood ratio was 4.4 times (2.2 to 8.8) and negative likelihood ratio of only 0.2 times (0.09 to 0.42). The best cut off valueof 0.2 was shown by the area under the curve of 83.5%. Based on this study, the diagnostic value of anti-dengue IgA had a good validityfor the diagnosis of dengue virus infection.
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Talarmin, Antoine, Bhety Labeau, Josiane Lelarge, and Jean-Louis Sarthou. "Immunoglobulin A-Specific Capture Enzyme-Linked Immunosorbent Assay for Diagnosis of Dengue Fever." Journal of Clinical Microbiology 36, no. 5 (1998): 1189–92. http://dx.doi.org/10.1128/jcm.36.5.1189-1192.1998.
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Dengue fever (DF) is usually diagnosed by testing for dengue virus immunoglobulin M (IgM) by a capture enzyme-linked immunosorbent assay (ELISA) (MAC-ELISA). However, IgM can last for months, and its presence might reflect a previous infection. We have tested the use of anti-dengue virus IgA capture ELISA (AAC-ELISA) for the diagnosis of DF by comparing the results of MAC-ELISAs and AAC-ELISAs for 178 serum samples taken from patients with confirmed cases of DF. IgM appears more rapidly (mean delay of positivity, 3.8 days after the onset of DF) than IgA (4.6 days) but lasts longer; the peak IgA titer is obtained on day 8. The specificity and the positive predictive value of AAC-ELISA are 100%; its sensitivity and negative predictive value (NPV) are also 100% between days 6 and 25 after the onset of DF, but they decrease drastically when data for tests conducted with specimens from the first days of infection are included, because the IgA titers, like the IgM titers, have not yet risen. AAC-ELISA is a simple method that can be performed together with MAC-ELISA and that can help in interprating DF serology.
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Akinshina, Yu A., V. F. Larichev, M. A. Saifullin, S. G. Mardanly, and A. M. Butenko. "COMPARISON OF THE APPLICATION OF DOMESTIC «ELISA-IGM-DENGUE» KIT, DELIVERED IN THE D.I. IVANOVSKY INSTITUTE OF VIROLOGY (MOSCOW, RUSSIAN FEDERATION) AND «ANTI-DENGUE VIRUS ELISA IGM» KIT (EUROIMMUN, GERMANY) FOR THE SERODIAGNOSIS OF DENGUE FEVER." Epidemiology and Infectious Diseases 22, no. 1 (February 15, 2017): 4–8. http://dx.doi.org/10.17816/eid40949.
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88 sera from patients with dengue fever were examined with the use of two test systems: Pilot Production kit «ELISA- IgM-dengue», delivered in the D.I. Ivanovsky Institute of Virology (Russian Federation) and. All 88 samples were positive for specific IgM in «ELISA-IgM-dengue», but 8 cases examined with the test kit «Anti-Dengue virus ELISA (IgM)» (Euroimmun, Germany) appeared to be negative. The ratio between the titers of anti-dengue IgM determined in patients’ blood samples by "ELISA-IgM-dengue" and «Ratio» (index, recommended by the company «Euroimmun» for differentiation of positive, equivocal or negative results, was evaluated. 53 blood sample containing M antibodies to cytomegalovirus (n = 43), Epstein-Barr virus (n = 6), herpes simplex virus (n = 2) and rubella virus (n = 2) were tested by both ELISA kits to compare their specificity. When «ELISA- IgM-dengue» kits were applied all observed samples were negative, but under the application of the «Anti-Dengue virus ELISA (IgM)» kit (Euroimmun, Germany) 17 samples (32.1%) were false positive or equivocal.
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Somlor, Somphavanh, Ludovic Brossault, and Marc Grandadam. "Evaluation of VIDAS® Diagnostic Assay Prototypes Detecting Dengue Virus NS1 Antigen and Anti-Dengue Virus IgM and IgG Antibodies." Diagnostics 11, no. 7 (July 7, 2021): 1228. http://dx.doi.org/10.3390/diagnostics11071228.
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Dengue is a serious tropical disease caused by the mosquito-borne dengue virus (DENV). Performant, rapid, and easy-to-use assays are needed for the accurate diagnosis of acute DENV infection. We evaluated the performance of three prototype assays developed for the VIDAS® automated platform to detect dengue NS1 antigen and anti-dengue IgM and IgG antibodies. Positive and negative agreement with competitor enzyme-linked immunosorbent assays (ELISA) and rapid diagnostic tests (RDT) was evaluated in 91 Lao patients (57 adults, 34 children) with acute DENV infection. The VIDAS® NS1 assay showed the best overall agreement (95.6%) with the competitor NS1 ELISA. Both VIDAS® NS1 and NS1 ELISA assays also demonstrated high sensitivity relative to DENV RNA RT-PCR set as gold standard (85.7% and 83.9%, respectively). In contrast, NS1 RDT was less sensitive relative to DENV RNA RT-PCR (72.7%). The overall agreement of VIDAS® IgM and IgG assays with the competitor assays was moderate (72.5% for IgM ELISA, 76.9% for IgG ELISA, and 68.7% for IgM and IgG RDT). In most analyses, test agreements of the VIDAS® assays were comparable in adults and children. Altogether, the VIDAS® dengue prototypes performed very well and appear to be suitable for routine detection of dengue NS1 antigen and anti-dengue IgM/IgG antibodies.
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Lima, Monique da Rocha Queiroz, Raquel Curtinhas de Lima, Elzinandes Leal de Azeredo, and Flavia Barreto dos Santos. "Analysis of a Routinely Used Commercial Anti-Chikungunya IgM ELISA Reveals Cross-Reactivities with Dengue in Brazil: A New Challenge for Differential Diagnosis?" Diagnostics 11, no. 5 (April 30, 2021): 819. http://dx.doi.org/10.3390/diagnostics11050819.
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In Brazil, chikungunya emerged in 2014, and by 2016, co-circulated with other arbovirosis, such as dengue and zika. ELISAs (Enzyme-Linked Immunosorbent Assays) are the most widely used approach for arboviruses diagnosis. However, some limitations include antibody cross reactivities when viruses belong to the same genus, and sensitivity variations in distinct epidemiological scenarios. As chikungunya virus (CHIKV) is an alphavirus, no serological cross reactivity with dengue virus (DENV) should be observed. Here, we evaluated a routinely used chikungunya commercial IgM (Immunoglobulin M) ELISA test (Anti-Chikungunya IgM ELISA, Euroimmun) to assess its performance in confirming chikungunya in a dengue endemic area. Samples (n = 340) representative of all four DENV serotypes, healthy individuals and controls were tested. The Anti-CHIKV IgM ELISA test had a sensitivity of 100% and a specificity of 25.3% due to the cross reactivities observed with dengue. In dengue acute cases, the chikungunya test showed an overall cross-reactivity of 31.6%, with a higher cross-reactivity with DENV-4. In dengue IgM positive cases, the assay showed a cross-reactivity of 46.7%. Serological diagnosis may be challenging and, despite the results observed here, more evaluations shall be performed. Because distinct arboviruses co-circulate in Brazil, reliable diagnostic tools are essential for disease surveillance and patient management.
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Galula, Jedhan Ucat, Gielenny M. Salem, Raul V. Destura, Roland Remenyi, and Day-Yu Chao. "Comparable Accuracies of Nonstructural Protein 1- and Envelope Protein-Based Enzyme-Linked Immunosorbent Assays in Detecting Anti-Dengue Immunoglobulin G Antibodies." Diagnostics 11, no. 5 (April 21, 2021): 741. http://dx.doi.org/10.3390/diagnostics11050741.
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Background: Dengue virus (DENV) infection remains a global public health concern. Enzyme-linked immunosorbent assays (ELISAs), which detect antibodies targeting the envelope (E) protein of DENV, serve as the front-line serological test for presumptive dengue diagnosis. Very few studies have determined the serostatus by detecting antibodies targeting the nonstructural protein 1 (NS1), which can function as diagnostic biomarkers to distinguish natural immunity from vaccine-induced immunity. Methods: We used community-acquired human serum specimens, with the serostatus confirmed by focus reduction microneutralization test (FRμNT), to evaluate the diagnostic performances of two NS1-based ELISA methods, namely, immunoglobulin G antibody-capture ELISA (NS1 GAC–ELISA) and indirect NS1 IgG ELISA, and compared the results with an E-based virus-like particle (VLP) GAC–ELISA. Results: NS1-based methods had comparable accuracies as VLP GAC–ELISA. Although the sensitivity in detecting anti-NS1 IgM was poor, indirect NS1 IgG ELISA showed similar limits of detection (~1–2 ng/mL) as NS1 GAC–ELISA in detecting anti-NS1 IgG. Combining the results from two or more tests as a composite reference standard can determine the DENV serostatus with a specificity reaching 100%. Conclusion: NS1-based ELISAs have comparable accuracies as VLP GAC–ELISA in determining dengue serostatus, which could effectively assist clinicians during assessments of vaccine eligibility.
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Lytton, Simon D., Mahmuda Yeasmin, Asish Kumar Ghosh, Md Rakibul Hassan Bulbul, Md Maruf Ahmed Molla, Martha Herr, Helmut Duchmann, et al. "Detection of Anti-Nucleocapsid Antibody in COVID-19 Patients in Bangladesh Is not Correlated with Previous Dengue Infection." Pathogens 10, no. 6 (May 22, 2021): 637. http://dx.doi.org/10.3390/pathogens10060637.
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Background: The assessment of antibody responses to severe acute respiratory syndrome coronavirus-2 is potentially confounded by exposures to flaviviruses. The aims of the present research were to determine whether anti-dengue antibodies affect the viral load and the detection of anti-coronavirus nucleocapsid (N)-protein antibodies in coronavirus infectious disease 2019 (COVID-19) in Bangladesh. Methods: Viral RNA was evaluated in swab specimens from 115 COVID-19 patients by real-time reverse transcription polymerase chain reaction (rT-PCR). The anti-N-protein antibodies, anti-dengue virus E-protein antibodies and the dengue non-structural protein-1 were determined in serum from 115 COVID-19 patients, 30 acute dengue fever pre-COVID-19 pandemic and nine normal controls by ELISA. Results: The concentrations of viral RNA in the nasopharyngeal; Ct median (95% CI); 22 (21.9–23.3) was significantly higher than viral RNA concentrations in oropharyngeal swabs; and 29 (27–30.5) p < 0.0001. Viral RNA concentrations were not correlated with-dengue IgG levels. The anti-nucleocapsid antibodies were IgA 27% positive and IgG 35% positive at days 1 to 8 post-onset of COVID-19 symptoms versus IgA 0% and IgG 0% in dengue patients, p < 0.0001. The levels of anti- nucleocapsid IgA or IgG versus the levels of anti-dengue IgM or IgG revealed no significant correlations. Conclusions: Viral RNA and anti-nucleocapsid antibodies were detected in COVID-19 patients from dengue-endemic regions of Bangladesh, independently of the dengue IgG levels.
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Chao, Day-Yu, Jedhan Ucat Galula, Wen-Fan Shen, Brent S. Davis, and Gwong-Jen J. Chang. "Nonstructural Protein 1-Specific Immunoglobulin M and G Antibody Capture Enzyme-Linked Immunosorbent Assays in Diagnosis of Flaviviral Infections in Humans." Journal of Clinical Microbiology 53, no. 2 (December 10, 2014): 557–66. http://dx.doi.org/10.1128/jcm.02735-14.
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IgM antibody- and IgG antibody-capture enzyme-linked immunosorbent assays (MAC/GAC-ELISAs) targeted at envelope protein (E) of dengue viruses (DENV), West Nile virus, and Japanese encephalitis virus (JEV) are widely used as serodiagnostic tests for presumptive confirmation of viral infection. Antibodies directed against the flavivirus nonstructural protein 1 (NS1) have been proposed as serological markers of natural infections among vaccinated populations. The aim of the current study is to optimize an IgM and IgG antibody-capture ELISA (MAC/GAC-ELISA) to detect anti-NS1 antibodies and compare it with anti-E MAC/GAC-ELISA. Plasmids to express premembrane/envelope (prM/E) or NS1 proteins of six medically important flaviviruses, including dengue viruses (DENV-1 to DENV-4), West Nile virus (WNV), and Japanese encephalitis virus (JEV), were constructed. These plasmids were used for the production of prM/E-containing virus-like particles (VLPs) and secreted NS1 (sNS1) from COS-1 cells. Archived clinical specimens from patients with confirmed DENV, JEV, and WNV infections, along with naive sera, were subjected to NS1-MAC/GAC-ELISAs before or after depletion of anti-prM/E antibodies by preabsorption with or without VLPs. Human serum specimens from previously confirmed DENV infections showed significantly enhanced positive-to-negative (P/N) ratios for NS1-MAC/GAC-ELISAs after the depletion of anti-prM/E antibodies. No statistical differences in sensitivities and specificities were found between the newly developed NS1- and VLP-MAC/GAC-ELISAs. Further application of the assays to WNV- and JEV-infected serum panels showed similar results. A novel approach to perform MAC/GAC-ELISAs for NS1 antibody detection was successfully developed with great potential to differentiate antibodies elicited by the tetravalent chimeric yellow fever-17D/dengue vaccine or DENV infection.
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Rochaeni, Ade, Aryati Aryati, Puspa Wardhani, and Usman Hadi. "ANALYSIS OF DENGUE SPECIFIC IMMUNE RESPONSE BASED ON SEROTYPE, TYPE AND SEVERITY OF DENGUE INFECTION (Analisis Respons Imun Spesifik Dengue terhadap Serotipe, Jenis dan Derajat Infeksi Virus Dengue)." INDONESIAN JOURNAL OF CLINICAL PATHOLOGY AND MEDICAL LABORATORY 23, no. 3 (April 14, 2018): 230. http://dx.doi.org/10.24293/ijcpml.v23i3.1199.
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Infeksi Virus Dengue (IVD) menimbulkan derajat klinis beragam dari DD hingga DBD/SSD. Respons imun spesifik dengue berupaIgM dan IgG anti dengue masih merupakan perdebatan untuk patogenesis DBD di samping faktor virulensi virus dan jenis infeksi.Penelitian ini bertujuan untuk menganalisis respons imun spesifik dengue terhadap serotipe, jenis dan derajat IVD di Surabaya. Subjekadalah pasien IVD yang dirawat di Ruang Tropik Infeksi Penyakit Dalam RSUD Dr. Soetomo dengan hasil penyaringan uji cepat NS1 (SDBioline Dengue Duo) dan/atau PCR (Simplexa Dengue) positif. Pemeriksaan IgM dan IgG anti dengue kuantitatif dengan metode ELISA(Panbio Dengue Duo IgM and IgG Capture). Penelitian dilakukan Maret–Agustus 2016 dan didapatkan 61 pasien dengan hasil NSI dan/atau PCR dengue positif. Identifikasi serotipe didominasi DEN-3, namun serotipe yang lebih virulen ditunjukkan DEN-1 yaitu semuapasien bermanifestasi sebagai infeksi sekunder dan DBD. Jenis infeksi primer sebanyak 19 (31,1%) dan infeksi sekunder 42 (68,9%).Derajat IVD meliputi DD 10 (16,4%), DBD 47 (77%) dan SSD 4 (6,56%). Nilai indeks rerata IgM dan IgG anti dengue di kelompokinfeksi serotipe DEN-1 (5,140 dan 5,774), DEN-2 (2,971 dan 2,222), DEN-3 (1,863 dan 2,792); kelompok jenis infeksi primer (1,478 dan0,746), sekunder (4,028 dan 4,864) dan kelompok derajat DD (1,170 dan 1,492), DBD I (3,370 dan 3,651), DBD II (3,924 dan 4,439)dan DBD III (4,164 dan 4,243). Sebagai simpulan respons imun spesifik dengue didapatkan lebih tinggi bermakna di kelompok infeksiserotipe DEN-1, kelompok jenis infeksi sekunder dan kelompok DBD/SSD.
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Vázquez, Susana, Gilda Lemos, Maritza Pupo, Oscar Ganzón, Daniel Palenzuela, Adriana Indart, and María G. Guzmán. "Diagnosis of Dengue Virus Infection by the Visual and Simple AuBioDOT Immunoglobulin M Capture System." Clinical Diagnostic Laboratory Immunology 10, no. 6 (November 2003): 1074–77. http://dx.doi.org/10.1128/cdli.10.6.1074-1077.2003.
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ABSTRACT The Dengue IgM Capture ELISA (MAC-ELISA) is the immunoenzymatic system recommended by the Pan American Health Organization and the World Health Organization for the serological diagnosis of dengue virus infection due to its high sensitivity, ease of performance, and use of a single acute-phase serum sample. However, tests with this enzyme-linked immunosorbent assay (ELISA) system are time-consuming and require equipment for washing, incubation, and reading of the results. AuBioDOT is a multistep visual diagnostic immunoassay that uses technology based on the immunoglobulin M (IgM) capture ELISA principle. This system uses white polyethylene opaque plates as the solid phase, colloidal gold as the marker, and silver ion amplification. It does not require special equipment, it is totally manually operated, and it can be performed in less than 1 h. The sensitivity and specificity of AuBioDOT for the detection of anti-dengue virus IgM antibodies were studied with a panel of 336 serum samples (150 serum samples from patients with suspected or serologically confirmed dengue virus infection, 186 serum samples from healthy blood donors and patients without dengue virus infection). The results were compared with those obtained by the MAC-ELISA. A sensitivity of 97.7% and a specificity of 97.1% were obtained. The concordance of the two tests was 97.3%, with a kappa index of 0.94. The application of AuBioDOT for the detection of anti-dengue virus IgM antibodies is recommended as an alternative method for the diagnosis of dengue virus infection, both for clinical diagnosis and for seroepidemiological surveillance. The system is useful under field conditions and in laboratories and requires little equipment.
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Perdomo-Celis, Federico, Doris Marta Salgado, and Carlos Fernando Narváez. "Desarrollo de una captura de Elisa para la detección de dengue-inmunoglobulina específica M." RFS Revista Facultad de Salud 8, no. 1 (January 2, 2016): 9. http://dx.doi.org/10.25054/rfs.v8i1.1317.
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El dengue es una importante enfermedad transmitida por vectores a nivel mundial. Su diagnóstico requiere confirmación por laboratorio. La detección de inmunoglobulina (Ig) M específica de virus dengue (DENV) por ensayo de inmunoadsorción ligado a enzima (ELISA) es un método frecuentemente usado. Sin embargo, las pruebas comerciales de ELISA de captura IgM para el diagnóstico de dengue son costosos, no están fácilmente disponibles y la entrega de resultados toma más del tiempo que el deseado en áreas endémicas. Aquí se propuso desarrollar una captura de ELISA para detección de IgM específica de DENV en plasma y ayudar al diagnóstico de dengue. El plasma de dos niños con infección aguda por DENV-2 confirmada y dos anticuerpos monoclonales anti-DENV hechos en ratón (uno serotipo-específico y el otro de reacción cruzada para los 4 serotipos) fueron evaluados. El ensayo fue efectivo en la detección de IgM específica de DENV y es comparable con un estuche comercialmente disponible, incluso a las mayores diluciones de la muestra y de los anticuerpos anti-DENV. Adicionalmente, en un niño con infección primaria por DENV-2, la IgM específica de DENV en plasma fue serotipo-específica. Este trabajo fortalece la capacidad tecnológica para el estudio y diagnóstico de la infección por DENV en un área endémica.
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Porter, Kevin R., Susana Widjaja, Handinata Darmawan, Lohita, Sri Hartati Hadiwijaya, Chairin Nisa Maroef, Wuryadi Suharyono, and Ratna Tan. "Evaluation of a Commercially Available Immunoglobulin M Capture Enzyme-Linked Immunosorbent Assay Kit for Diagnosing Acute Dengue Infections." Clinical Diagnostic Laboratory Immunology 6, no. 5 (September 1, 1999): 741–44. http://dx.doi.org/10.1128/cdli.6.5.741-744.1999.
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ABSTRACT Recently, commercially available kits for the detection of anti-dengue virus (anti-DEN) immunoglobulin M (IgM) antibodies have been developed. These standardized assays have greatly enhanced our ability to effectively diagnose DEN infections. We conducted an evaluation of a test kit manufactured by MRL Diagnostics Inc. that is designed to detect anti-DEN IgM antibodies. Eighty paired samples from DEN-infected individuals were tested by the MRL DEN Fever Virus IgM Capture enzyme-linked immunosorbent assay (ELISA), the PanBio Duo ELISA, the PanBio Rapid Immunochromatographic Test (PRIT), and the IgM-IgG antibody capture (MAC/GAC) ELISA. All infections were confirmed by either PCR-assisted detection of DEN transcripts or by DEN isolation in C6/36 cells. Seventeen paired samples from individuals with no evidence of acute DEN infection were used as negative controls. The PRIT had the best sensitivity (100%), whereas the MAC/GAC ELISA and the PanBio Duo assay had the highest levels of specificity. The MRL ELISA and the PanBio Duo assay were the top performers when taking into consideration both sensitivity and specificity. All assays were able to detect DEN-specific antibodies in samples from patients with either primary or secondary infections, regardless of the infecting DEN serotype.
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Manuel, Samantha, Liane Virginia-Cova, Loubiela Joseph, Chris Roggeveen, and Radjin Steingrover. "Zika Virus Serologic Diagnosis by NS1 ELISA in Curacao." Open Forum Infectious Diseases 4, suppl_1 (2017): S302—S303. http://dx.doi.org/10.1093/ofid/ofx163.698.
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Abstract Background Zika virus (ZIKV) was introduced in the Caribbean island of Curacao in January 2016. A commercially available ZIKV IgM and IgG ELISA was evaluated on patients that were PCR-positive for ZIKV. Methods ZIKV infection was established by PCR in urine samples. Samples from PCR-positive patients were selected for validation of a ZIKV NS1 IgG and IgM ELISA. Patients with a follow-up sample ≥ 2 weeks after initial presentation were used to assess the sensitivity of the assay. Samples of 15 historical controls with serological evidence of Dengue, Chikungunya or an unrelated viral infection were included to establish specificity and cross-reactivity. Results Fourteen patients with positive ZIKV PCR diagnosis had repeated serum samples drawn ≥ 2 weeks after the initial sample. The combined results of these repeated IgM and IgG tests resulted in a sensitivity of 92%. One pregnant female showed no presence of IgG or IgM in any of the two samples. Testing of the panel of historical ZIKV-negative controls resulted in a specificity of 100% in both the quantitative and semi-quantitative setting of the ELISA. One patient with known high-titers of antibodies against Chikungunya virus in the respective panel displayed borderline reactive results for ZIKV IgG in both quantitative and semi-quantitative setting of the assay. Conclusion In this PCR-positive ZIKV cohort of patients, the newly available ZIKV NS1 ELISA displayed excellent performance characteristics. Cross-reactivity was indicated for Chikungunya in one case. No cross-reactivity was found for Dengue virus infection. One pregnant female showed no signs of developing anti-ZIKV IgM or IgG in this study. In the light of intrauterine pathogenesis, the lack of development of maternal IgG during ZIKV infection is a concern. Disclosures All authors: No reported disclosures.
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Retno Setyowati, Ety, Aryati Aryati, Prihatini Prihatini, and M. Y. Probohoesodo. "EVALUASI PEMERIKSAAN IMUNOKROMATOGRAFI UNTUK MENDETEKSI ANTIBODI IgM DAN IgG DEMAM BERDARAH DENGUE ANAK." INDONESIAN JOURNAL OF CLINICAL PATHOLOGY AND MEDICAL LABORATORY 12, no. 2 (March 14, 2018): 88. http://dx.doi.org/10.24293/ijcpml.v12i2.850.
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The gold standard diagnosis of DHF by RT-PCR needs a complex technology and is time consuming. Serological tests have beendeveloped to detect IgM and IgG anti dengue to determine primary as well as secondary acute phase infection. IgM and IgG antidenguetests by immunochromatography have been used, due to a high diagnostic validity, also because they are simple, practicable, easy, rapid(15–30 minutes), can be used in a single serum sample. ELISA method has been used as a confirmation method. The aim of this studyis to evaluate the immunochromatography method in detecting IgG and IgM anti dengue of DHF patients. The study was performedon 50 serum samples from patients of the ICU Department of Paediatrics Dr. Soetomo Hospital, Surabaya during July–August 2005with dengue virus infection according to the 1997, WHO criterion and 27 serum samples from non dengue virus infection patients.ELISA method showed positive infection in 44 samples. Immunochromatography method showed positive infection in 43 samples, butwas negative in 1 sample. Diagnostic sensitivity of Immunochromatography is 97.7% (43/44) and the diagnostic specificity is 92.6%(25/27). Immunochromatography method has a high diagnostic value in assisting the diagnosis of DHF.
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Sharma, Mukesh. "Role of Pro-inflammatory IL-8 and Anti-inflammatory IL-10 Cytokines in Dengue Severity." Journal of Communicable Diseases 53, no. 02 (June 30, 2021): 69–75. http://dx.doi.org/10.24321/0019.5138.202128.
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Introduction: During dengue infection, cytokine levels may increase as various cytokines are released from infected inflammatory cells. This study was conducted to measure the levels of cytokines IL-8 and IL-10 in dengue patients and correlate them with dengue severity. Material & Methods: A prospective study was conducted on febrile patients suspected of dengue fever, seeking medical care in our institute. 107 cases confirmed to have dengue fever (by NS1/ IgM ELISA) and 100 healthy individuals with age and sex matched, were included in the study. The clinical features of all patients were recorded, and cytokine levels of IL-8 and IL-10 were estimated by ELISA in the dengue patients and healthy controls. Results: Out of 400 febrile patients suspected of having dengue fever, 107 (26.75%) cases were confirmed cases, of which 56 (52.3%), 20 (18.7%), and 31 (29%) were positive for only NS1 antigen, only IgM antibody, and both NS1 and IgM, respectively. Depending on the severity of the disease, 9 (8.5%) cases were classified as severe dengue cases while 98 (91.5%) as non-severe dengue fever. Mean levels (pg/ml) for IL-8 were 281.6 ± 76.6, 150.41 ± 55.9 and 75.4 ± 49.2 in severe dengue, dengue fever, and healthy controls respectively while for IL-10, the values were 219.4 ± 150.5, 38.9 ± 67.2, and 6.6 ± 0.65 among severe dengue cases, dengue cases, and healthy controls, respectively. Conclusion: Mean level of cytokines IL-8 and IL-10 were significantly raised in severe dengue patients as compared to non-severe dengue patients and healthy controls, suggesting their role in causing severe disease and as a potential predictor for disease severity and fatal outcome.
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Rantam, Fedik. "Serotype Infectivity and Phylogenetic of Dengue Virus cause of Dengue Fever (DF), Dengue Hemorrhagic Fever (DHF), and Dengue Shock Syndrome (DSS) in Surabaya- Indonesia." Journal of Stem Cell Research and Tissue Engineering 4, no. 1 (August 26, 2020): 1. http://dx.doi.org/10.20473/jscrte.v4i1.21588.
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Infection with DENV causes a spectrum of clinical disease ranging. The aim of this study is to investigate the infectivity of DENV with degree of severity dengue infection in Surabaya. Dengue infection was established by IgM anti dengue, and two step multiplex RT PCR and Nucleotide sequence. Grading of degree severity infection follow the WHO criteria 2011. DSS cases found 3 from 36 patients caused by DENV 2. The most uninfective was DENV 1, and the most prevalence dengue infection caused by DENV 3. The infectivity of dengue infection shown 16 patients lead to severity with plasma leakage. All of sera patients detecting using multiplex RT-PCR were positive, but it were analyzed using Duo ELISA only 22 serum sera positive IgM and IgG from 36 sera. . The Phylogenetic analysis indicates that the isolates from 2011 to 2012 close related with dengue isolate from 1998 and belong to 2009 to 2020.In this study it indicates that DENV 2 predominantly is the cause of DSS.
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Lu, Yixiao, Onanong Sengvilaipaseuth, Anisone Chanthongthip, Ooyanong Phonemixay, Manivanh Vongsouvath, Phonelavanh Phouminh, Stuart D. Blacksell, Paul N. Newton, and Audrey Dubot-Pérès. "Comparison of Two Commercial ELISA Kits for the Detection of Anti-Dengue IgM for Routine Dengue Diagnosis in Laos." Tropical Medicine and Infectious Disease 4, no. 3 (July 25, 2019): 111. http://dx.doi.org/10.3390/tropicalmed4030111.
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The endemicity of Dengue virus (DENV) infection remains a major public health problem in Lao PDR. In this study, we compared two commercial anti-dengue IgM ELISA kits, Panbio® Dengue IgM Capture ELISA (Panbio Kit, Alere, Waltham, MA, USA) and DEN DetectTM MAC-ELISA (InBios kit, InBios International, Inc., Seattle, WA, USA), in the context of diagnosis of patients admitted to hospital with clinical dengue presentation. Two panels of paired blood samples were tested. Panel A was composed of 54 dengue confirmed patients (by DENV real-time RT-PCR) and 11 non-dengue dengue patients (other infections confirmed by corresponding PCR results). Panel B included 74 patients randomly selected from consecutive patients admitted to Mahosot Hospital in 2008 with suspicion of dengue fever according to WHO criteria. Results from panel A showed significantly better sensitivity for Panbio kit (64.8%; 95%CI: 50.6–77.3%) than for InBios kit (18.5%; 95%CI: 9.3–31.4%) when testing admission sera. Sensitivity was increased for both kits when combining results from admission and convalescent sera. Concordant results were obtained from panel B with fair agreement (κ = 0.29) between both kits when testing single admission samples, and moderate agreement (κ = 0.5) when combining results from admission and convalescent sera.
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Ambrose, Jason H., Shamala Devi Sekaran, and Azliyati Azizan. "Dengue Virus NS1 Protein as a Diagnostic Marker: Commercially Available ELISA and Comparison to qRT-PCR and Serological Diagnostic Assays Currently Used by the State of Florida." Journal of Tropical Medicine 2017 (2017): 1–6. http://dx.doi.org/10.1155/2017/8072491.
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Background. The proper management of patients infected with dengue virus requires early detection. Here, real-time molecular assays have proven useful but have limitations, whereas ELISAs that detect antibodies are still favored but results are obtained too late to be of clinical value. The production of DENV NS1 peaks early during infection and its detection can combine the advantages of both diagnostic approaches.Methods.This study compared assays currently used for detecting DENV infection at the Florida Department of Health including anti-DENV IgM and IgG ELISAs as well as qRT-PCR, against a commercially available DENV NS1 ELISA. These comparisons were made among a group of 21 human sera.Results. Nine of 14 (64.3%) DENV qRT-PCR+ samples were also DENV NS1+. Interestingly, the 5 NS1− samples that were qRT-PCR+ were additionally IgM− and IgG+ suggesting a nonprimary infection. Compared to qRT-PCR, the NS1 assay had a sensitivity of 64.3%, specificity 100%, PPV of 100%, and NPV of 58.3%.Conclusions. The NS1 ELISA performed as expected in known DENV qRT-PCR+ samples; however negative NS1 results for qRT-PCR+ and IgG+ sera seemingly reduced the usefulness of the NS1 ELISA for nonprimary cases. We therefore conclude that diagnosis obtained via DENV NS1 ELISA deserves further investigation.
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VASCONCELOS, Pedro F. C., Amélia P. A. TRAVASSOS DA ROSA, Ivo C. B. COELHO, Dalgimar B. MENEZES, Elizabeth S. TRAVASSOS DA ROSA, Sueli G. RODRIGUES, and Jorge F. S. TRAVASSOS DA ROSA. "Involvement of the central nervous system in dengue fever: three serologically confirmed cases from Fortaleza, Ceará, Brazil." Revista do Instituto de Medicina Tropical de São Paulo 40, no. 1 (January 1998): 35–40. http://dx.doi.org/10.1590/s0036-46651998000100008.
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Three cases of dengue fever involving the central nervous system (CNS) are reported. All occurred in 1994 during a dengue (DEN) epidemic caused by serotypes DEN-1 and DEN-2. The first case examined was a 17-year-old girl who complained of fever, nuchal rigidity and genital bleeding. Three blood samples were positive by anti-dengue IgM ELISA and showed hemagglutination-inhibition (HI) test titers <FONT FACE="Symbol">³</font> 1,280. The second case concerned a 86-year-old woman with fever, muscle and joint pains, altered consciousness, syncope, nuchal rigidity and meningismus. Her blood sample showed an HI titer of 1:320 for flaviviruses, and an IgM ELISA positive for dengue. The third case was a 67-year-old woman with fever, abnormal behaviour, seizures, tremor of extremities, thrombocytopenia, increased hematocrit and leukopenia. The patient suffered a typical case of dengue hemorrhagic fever with ensuing shock and a fatal outcome. A single blood sample showed HI antibodies of <FONT FACE="Symbol">³</font> 1,280 and an IgM ELISA positive for dengue. No virus could be isolated from any patient by inoculation of blood into C6/36 cells and suckling mice. No other agent of disease was encountered in the patient.
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Narkwa, P. W., M. Mutocheluh, T. B. Kwofie, M. Owusu, A. Annan, I. Ali, and J. K. Boamah. "Dengue virus exposure among blood donors in Ghana." Journal of Medical and Biomedical Sciences 5, no. 2 (October 20, 2016): 30–35. http://dx.doi.org/10.4314/jmbs.v5i2.5.
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Dengue is an urban arbovirus whose aetiologic agent is the flavivirus with four distinct antigen serotypes (DENV-1, DENV-2, DENV-3 and DENV-4) that is transmitted to humans through the bite of the mosquito Aedes aegypti. Ghana is endemic for Aedes aegypti mosquitoes and probably dengue viruses. Due to limited data on dengue virus exposure among Ghanaians, we surveyed 188 healthy adult blood donors for the presence of IgG and IgM antibodies to the four serotypes of dengue. Five milliliters of peripheral blood from the blood donors were collected in plain tubes. Serum was then obtained and ELISA tests were employed to detect both dengue virus total antibodies and IgM. The samples were further tested for dengue virus RNA using RT-PCR. Dengue virus IgG was positive for 43.6% of all the 188 blood donor samples tested but all donors were negative for anti-dengue IgM antibody and dengue virus RNA. The rate of dengue virus total antibody exposure did not differ statistically between urban and rural districts. This study shows for the first time that some regions of Ghana are hyperendemic for dengue virus infection but suggests blood for transfusion is invariably dengue virus free. This report has provided a baseline data that will inform wider discussions about the impact of this dengue fever and also guide policy makers to develop effective and affordable early warning and outbreak response systems for Ghana.Journal of Medical and Biomedical Sciences (2016) 5(2), 30-35
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Rojas, Alejandra, Fátima Cardozo, César Cantero, Victoria Stittleburg, Sanny López, Cynthia Bernal, Francisco Eugenio Gimenez Acosta, et al. "Characterization of dengue cases among patients with an acute illness, Central Department, Paraguay." PeerJ 7 (October 9, 2019): e7852. http://dx.doi.org/10.7717/peerj.7852.
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Background In 2018, Paraguay experienced a large dengue virus (DENV) outbreak. The primary objective of this study was to characterize dengue cases in the Central Department, where the majority of cases occur, and identify factors associated with DENV infection. Methods Patients were enrolled from January-May 2018 if they presented with a suspected arboviral illness. Acute-phase specimens (≤8 days after symptom onset) were tested using rRT-PCR, a rapid diagnostic test for DENV nonstructural protein 1 (NS1) and anti-DENV IgM and IgG, and ELISA for IgG against NS1 from Zika virus (ZIKV). Results A total of 231 patients were enrolled (95.2% adults) at two sites: emergency care and an outpatient clinical site. Patients included 119 (51.5%) dengue cases confirmed by rRT-PCR (n = 115, 96.6%) and/or the detection of NS1 and anti-DENV IgM (n = 4, 3.4%). DENV-1 was the predominant serotype (109/115, 94.8%). Epidemiologically, dengue cases and non-dengue cases were similar, though dengue cases were less likely to reside in a house/apartment or report a previous dengue case. Clinical and laboratory findings associated with dengue included red eyes, absence of sore throat, leucopenia and thrombocytopenia. At an emergency care site, 26% of dengue cases (26/100) required hospitalization. In univariate analysis, hospitalization was associated with increased viral load, anti-DENV IgG, and thrombocytopenia. Among dengue cases that tested positive for IgG against ZIKV NS1, the odds of DENV NS1 detection in the acute phase were decreased 10-fold (OR 0.1, 0.0–0.3). Conclusions Findings from a predominantly adult population demonstrate clinical and laboratory factors associated with DENV infections and the potential severity of dengue in this group. The combination of viral load and specific IgG antibodies warrant further study as a prognostic to identify patients at risk for severe disease.
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Shah, Y., G. Khadka, GP Gupta, N. Adhikari, A. Poudel, KP Pant, B. Dahal, and BD Pandey. "Sero-diagnosis of Dengue virus in Different Hospitals of Nepal." International Journal of Infection and Microbiology 1, no. 2 (January 20, 2013): 58–62. http://dx.doi.org/10.3126/ijim.v1i2.7003.
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INTRODUCTION: Dengue fever (DF) is an emerging mosquito borne viral disease and important public health problem in low land Terai region which is also moving towards hilly region Nepal. This study was designed to determine the sero-prevalence of dengue virus infection in patients visiting hospitals of Nepal. MATERIALS AND METHODS: This study was conducted during period (June-November) of 2010 in Nepalese patients with fever visiting hospitals of Birganj, Damouli, Biratanagar, Dhading Besi and Chitwan. The sero-prevalence of dengue virus specific IgM was determined by enzyme linked immunosorbent assay (ELISA). Serum samples were collected from 289 patients visiting hospitals with history of fever and clinically suspected dengue fever. RESULTS: The anti-dengue IgM positivity was found to be 8.99%. The positive dengue cases were higher in male (10.8%) as compared to female (7.1%) though it was not statistically significant (P>0.05). Among different age groups, the highest positive cases (12.3%) were from age group below 15 years followed by above 50 years 8.3%. Out of 5 hospitals, the highest positive cases were in Tanahu hospital, Damouli (23.8%) followed by Bharatpur hospital and Chitwan (22.2%). Age and gender were found to be independent predictors. The highest numbers of dengue positive cases were in occupation group business (13.3%) followed by agriculture (12.7%). CONCLUSIONS: Prevalence of dengue virus infection is increasing and proper control measure should be provided. IgM capture ELISA was used for laboratory analysis and remains as a reliable and inexpensive method for the diagnosis of dengue. Hence, the IgM capture ELISA has become the most accepted technique for the diagnosis of dengue in developing countries like Nepal. DOI: http://dx.doi.org/10.3126/ijim.v1i2.7003 Int J Infect Microbiol 2012;1(1):58-62
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Granger, Dane, Heather Hilgart, Lori Misner, Jaime Christensen, Sarah Bistodeau, Jennifer Palm, Anna K. Strain, et al. "Serologic Testing for Zika Virus: Comparison of Three Zika Virus IgM-Screening Enzyme-Linked Immunosorbent Assays and Initial Laboratory Experiences." Journal of Clinical Microbiology 55, no. 7 (April 26, 2017): 2127–36. http://dx.doi.org/10.1128/jcm.00580-17.
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ABSTRACT Serologic evaluation for Zika virus (ZIKV) infection currently includes an initial screen using an anti-ZIKV IgM antibody capture enzyme-linked immunosorbent assay (MAC-ELISA) followed by supplemental testing of specimens with nonnegative results by a plaque reduction neutralization test (PRNT). We compared the performance characteristics of three ELISAs for the detection of IgM class antibodies to ZIKV, including the Centers for Disease Control and Prevention (CDC) Zika MAC-ELISA, the InBios ZIKV Detect MAC-ELISA, and the Euroimmun anti-Zika Virus IgM ELISA. Additionally, we present our initial experiences with ZIKV serologic testing from a national reference laboratory perspective. Using both retrospectively and prospectively collected specimens from patients with possible ZIKV infection, we show that the CDC and InBios MAC-ELISAs perform comparably to each other, with positive agreement, negative agreement, and interrater kappa values ranging from 87.5% to 93.1%, 95.7% to 98.5%, and 0.52 to 0.83, respectively. In contrast, comparison of the Euroimmun ZIKV ELISA to either the CDC or InBios MAC-ELISAs resulted in positive agreement, negative agreement, and interrater kappa values ranging from 17.9% to 42.9%, 91.7% to 98.6%, and 0.10 to 0.39, respectively. Among the 19 prospective samples submitted for PRNT, nine were negative, eight specimens had neutralizing antibodies to a flavivirus (unable to be identified), and one sample each was confirmed for ZIKV or dengue virus infection. This study highlights the ongoing challenges associated with serologic diagnosis of ZIKV infection. Although the availability of a commercial serologic test for ZIKV has greatly expanded the national capacity for such testing, the need to further characterize and improve these assays, particularly with regard to specificity, remains.
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Raza, Faiz Ahmed, Hasnain Javed, Muhammad Mujeeb Khan, Obaid Ullah, Areeba Fatima, Muhammad Zaheer, Saima Mohsin, Shahida Hasnain, Ruqyya Khalid, and Arslan Ahmed Salam. "Dengue and Chikungunya virus co-infection in major metropolitan cities of provinces of Punjab and Khyber Pakhtunkhwa: A multi-center study." PLOS Neglected Tropical Diseases 15, no. 9 (September 23, 2021): e0009802. http://dx.doi.org/10.1371/journal.pntd.0009802.
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Dengue has become endemic in Pakistan with annual recurrence. A sudden increase in the dengue cases was reported from Rawalpindi in 2016, while an outbreak occurred for the first time in Peshawar in 2017. Therefore, a multi-center study was carried out to determine the circulating dengue virus (DENV) serotypes and Chikungunya virus (CHIKV) co-infection in Lahore, Rawalpindi, and Peshawar cities in 2016–18. A hospital-based cross-sectional study was carried out in Lahore and Rawalpindi in 2016–18, while a community-based study was carried out in Peshawar in 2017. The study participants were tested for dengue NS1 antigen using an immunochromatographic device while anti-dengue IgM/IgG antibodies were detected by indirect ELISA. All NS1 positive samples were used for DENV serotyping using multiplex real-time PCR assay. Additionally, dengue samples were tested for CHIKV co-infection using IgM/IgG ELISA. A total of 6291 samples were collected among which 8.11% were NS1 positive while 2.5% were PCR positive. DENV-2 was the most common serotype (75.5%) detected, followed by DENV-1 in 16.1%, DENV-3 in 3.9% and DENV-4 in 0.7% while DENV-1 and DENV-4 concurrent infections were detected in 3.9% samples. DENV-1 was the predominant serotype (62.5%) detected from Lahore and Rawalpindi, while DENV-2 was the only serotype detected from Peshawar. Comorbidities resulted in a significant increase (p-value<0.001) in the duration of hospital stay of the patients. Type 2 diabetes mellitus substantially (p-value = 0.004) contributed to the severity of the disease. Among a total of 590 dengue positive samples, 11.8% were also positive for CHIKV co-infection. Co-circulation of multiple DENV serotypes and CHIKV infection in Pakistan is a worrisome situation demanding the urgent attention of the public health experts to strengthen vector surveillance.
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Bhat, Vivek G., Preeti Chavan, Shashank Ojha, and Pravin K. Nair. "Challenges in the Laboratory Diagnosis and Management of Dengue Infections." Open Microbiology Journal 9, no. 1 (July 31, 2015): 33–37. http://dx.doi.org/10.2174/1874285801509010033.
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Dengue fever is considered the most important arthropod-borne viral diseases in terms of morbidity and mortality. An accurate and efficient diagnosis of dengue plays an important role in case confirmation. The virus may be isolated during the viremic phase (within day 5 of illness), from serum, plasma and peripheral blood mononuclear cells. Enzyme linked immunoassay (ELISA) has demonstrated the presence of high levels of dengue NS1 antigen and tests may be performed by enzyme-immunoassays (EIAs) or immune-chromatographic (ICT) methods. These assays are specific with respect to different flaviviruses. Conventional and real time RT PCR, nested PCR, multiplex PCR and Nucleic acid sequence based amplification (NASBA) have been described as sensitive and relatively rapid method of detecting the virus during the early viremic phase. Other tests used include assay of anti-dengue specific IgM and IgG ELISA. Currently no curative treatment in terms of anti-viral drugs is available for dengue and patients are managed with rest and aggressive supportive therapy. Management may be done at home or in the hospital depending on the severity of the illness. Hospital management includes fluid therapy, blood component transfusion and other modalities of treatments like steroids, recombinant factor VII and management of complications. Various vaccines are in trial stages and may become available in the near future.
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Parkash, Om, Muhammad Amiruddin Abdullah, Chan Yean Yean, Shamala Devi Sekaran, and Rafidah Hanim Shueb. "Development and Evaluation of an Electrochemical Biosensor for Detection of Dengue-Specific IgM Antibody in Serum Samples." Diagnostics 11, no. 1 (December 26, 2020): 33. http://dx.doi.org/10.3390/diagnostics11010033.
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Dengue is an arbovirus disease transmitted mainly by Aedes mosquitoes. As dengue shares similar clinical symptoms with other infectious diseases, prompt and accurate diagnosis is pivotal to clinicians’ decisions on appropriate management. Conventional diagnostic tests to detect the dengue-specific IgM antibody are limited in their performance and ease of use. To address these issues, we developed and evaluated a biosensor based on screen-printed carbon electrodes (SPCEs) for the detection of dengue-specific immunoglobulin M (IgM) antibodies. Various optimisations were performed in order to increase the sensitivity and specificity of the biosensor. For optimal and proper orientation of the paratope sites of goat anti-human IgM capture antibodies (GAHICA), various antibody techniques, including passive, covalent, protein A, protein G and streptavidin/biotin systems, were tested on the SPCEs. The assay reagents for the biosensor were also optimised prior to its evaluation. Analytical sensitivity evaluation was carried out using pooled sera, while analytical specificity evaluation was conducted on a panel of six non-dengue serum samples. Subsequently, diagnostic sensitivity and specificity evaluation were performed using 144 reference samples. Electrochemical current signals generated from H2O2 catalysed by HRP-labelled anti-dengue detection antibodies were measured using the chronoamperometric technique. With a limit of detection (LOD) of 106 serum dilution, the analytical sensitivity of the developed biosensor was 10 times higher than commercial ELISA. The analytical specificity of this dengue IgM biosensor was 100%. Similarly, the biosensor’s diagnostic performance was 100% for sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV). These findings suggest that the developed biosensor has a great potential to be used to diagnose dengue after seroconversion.
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Shah, Yogendra, Govind Prasad Gupta, Kishor Pandey, Sher Bahadur Pun, Krishna Prasad Pant, Santosh Dhakal, and Basu Dev Pandey. "Dengue Virus Detection by Serological and Molecular Method in Different Hospitals of Nepal." Medical Journal of Shree Birendra Hospital 11, no. 2 (April 5, 2013): 24–28. http://dx.doi.org/10.3126/mjsbh.v11i2.7905.
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Introduction: Dengue is an emerging mosquito-borne viral disease in the world and is the serious public health problem of Nepal. Methods: This study was designed to determine sero-epidemiology of dengue virus infection during the period (June-Nov) of 2010 among suspected patients with fever visiting Koshi Zonal Hospital (KZH), Biratnagar, Narayani sub-regional Hospital (NSH), Birgunj, Sukraraj Tropical and Infectious Disease Hospital (STIDH), Kathmandu and Dhading District Hospital (DDH), Dhadingbeshi. The sero-prevalence of anti-dengue IgM antibody was determined by enzyme linked immunosorbent assay (ELISA). Results: Among 271 serum samples tested, the anti-dengue IgM positivity was 14.4%. Sero-positivity in male was 10.7% of total and that in female was 3.7%. Among different age groups, the highest positive cases 11.8% were from age group 15-50 years and found least among the age group above 50 years 0.4%. Out of 4 different hospitals, the highest positive positive cases from STIDH with 9.2% and the least positive cases were from DDH (0.4%). RT-PCR showed 4.7% positivity of 21 samples tested. Conclusions: Enzyme immunoassay and RT-PCR serological marker can be used to diagnose the acute patients of dengue during outbreaks.Medical Journal of Shree Birendra Hospital; July-December 2012/vol.11/Issue2/24-27 DOI: http://dx.doi.org/10.3126/mjsbh.v11i2.7905
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Abhyankar, A. V., P. K. Dash, P. Saxena, R. Bhargava, M. M. Parida, A. M. Jana, A. K. Sahni, and P. V. L. Rao. "Comparison of a Dipstick Dot-ELISA with Commercial Assays for Anti-Dengue Virus IgM Antibodies." Viral Immunology 19, no. 4 (December 2006): 630–36. http://dx.doi.org/10.1089/vim.2006.19.630.
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Gupta, B. P., S. K. Mishra, K. D. Manandhar, R. Malla, C. S. Tamarakar, P. P. Raut, S. K. Sah, S. Pokhrel, R. Rauniyar, and A. Bajaracharya. "Seroprevalence 0f Dengue Virus Infection in Nepal." International Journal of Applied Sciences and Biotechnology 1, no. 4 (December 21, 2013): 224–27. http://dx.doi.org/10.3126/ijasbt.v1i4.9135.
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Dengue Virus infection is an emerging mosquito-borne disease. It is a global health problem and its expanding endemicity towards new territories is a serious concern. Relatively a new disease in Nepalese context, dengue abruptly appeared as massive outbreak in 2010, merely four years after its first introduction. It is a nagging public health problem in the low lands of Terai, expanding to new areas of Nepal in recent years. A cross-sectional study was conducted to determine anti-Dengue IgM positive rate in Lumbini, Dhading and Chitwan district. The study was carried from June 2012 to November 2012. The total number of Serum samples was collected from 275 patients visiting hospitals with history of fever, headache and suspected DF. The samples were examined by ELISA. The anti-Dengue IgM positivity was found to be 29.09 %. The positive rate was highest in Dhading (70.37%) followed by Bharatpur (37.6%) and Lumbini (11.38%). The Dengue positive cases were higher in males (32.5 %) than female (24.8 %). The highest positive cases (41.6%) were from age group less than 15 years. Dengue has substantial expansion in Western and Far Western Terai region of Nepal which was limited to the middle Terai region in the past and mostly infects older people.DOI: http://dx.doi.org/10.3126/ijasbt.v1i4.9135 Int J Appl Sci Biotechnol, Vol. 1(4): 224-227
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Johnson, Alison J., Denise A. Martin, Nick Karabatsos, and John T. Roehrig. "Detection of Anti-Arboviral Immunoglobulin G by Using a Monoclonal Antibody-Based Capture Enzyme-Linked Immunosorbent Assay." Journal of Clinical Microbiology 38, no. 5 (2000): 1827–31. http://dx.doi.org/10.1128/jcm.38.5.1827-1831.2000.
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Monoclonal antibody (MAb)-based capture enzyme-linked immunosorbent assays (ELISAs) for the detection of anti-arboviral immunoglobulin G (IgG ELISAs) were developed for a comprehensive array of medically important arboviruses from the Alphavirus,Flavivirus, and Bunyavirus genera. Tests were optimized and standardized so that maximum homology could be maintained among working parameters for the different viral agents, enabling a wide range of viruses to be easily tested for at one time. MAbs were screened for suitability as capture vehicles for antigens from the three genera. The final test configuration utilized group-reactive MAbs eastern equine encephalitis virus 1A4B-6, dengue 2 virus 4G2, and La Crosse encephalitis virus 10G5.4 to capture the specific inactivated viral antigens. Serum IgG was detected by using alkaline phosphatase-conjugated anti-human IgG (Fc portion). A dilution of 1:400 was chosen as the universal screening serum dilution, with endpoint titrations of serum samples testing positive eliminating occasional false-positive results. IgG ELISA results correlated with those of the standard plaque-reduction neutralization assays. As expected, some test cross-reactivity was encountered within the individual genera, and tests were interpreted within the context of these reactions. The tests were standardized for laboratory diagnosis of arboviral infections, with the intent that they be used in tandem with the corresponding IgM antibody-capture ELISAs.
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Mili, N. Y., S. Sikder, A. Basar, and E. Hoque. "Biochemical Profile and Degree of Liver Involvement in Dengue Fever." Journal of Medical Science & Research 27, Number 2 (July 1, 2017): 3–6. http://dx.doi.org/10.47648/jmsr.2017.v2702.01.
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Dengue disease has emerged globally as the most frequent and medically relevant viral infection transmitted by mosquito bite. Acute hepatitis is a manifestation of dengue virus infection. This study shows the impact of dengue on liver function was studied by biochemical tests on 80 patients out of them 53 male (age 42±12 yrs) and 27 female (age 39±13 yrs). The patients were diagnosed as dengue fever and were admitted in Holy Family Red Crescent Medical college Hospital from June 2014 to December 2016. All the patient were diagnosed by anti-dengue IgM positive by ELISA method. Abnormal level of aspartate aminotransferase (ASV, alanine aminotransferase (ALT), bilirubin, alkaline Phosphatase (Alp), gamma-glutamyl transferase (G-GT) and albumin and urinary albumin were observed in 82.5%, 82.5%, 47.5%, 38.75%, 71.25% ,66.25% and, 76.25% of the patients respectively. It is concluded that dengue fever may cause hepatic injury and transaminase elevation similar to that in patients with conventional viral hepatitis. In epidemic or endemic areas, dengue fever should be considered in the differential diagnosis of acute hepatitis.
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Gupta, GP, Y. Shah, A. Poudel, R. Pun, KP Pant, R. Kshetri, K. Pandey, and BD Pandey. "Serological and Molecular Study of Dengue Viruses in Different Hospitals of Nepal." Nepal Journal of Medical Sciences 2, no. 1 (February 21, 2013): 20–25. http://dx.doi.org/10.3126/njms.v2i1.7646.
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Background: Dengue Virus (DV) is an emerging mosquito borne viral disease and important public health problem in low land of Terai region which is also expanding to hilly region. Methods: This study was designed to estimate sero-prevalence of dengue virus infection in the post monsoon period (Jun-Dec) of 2010 in Nepalese patients with fever visiting hospitals of Birganj, Damouli, Biratnagar and Dhading Besi. Serum samples were collected from 280 patients visiting hospitals with history of fever & clinically suspected dengue fever. The sero-prevalence of dengue virus specific IgM was determined by enzyme linked immunosorbent assay (ELISA) kit (SD, Korea) Results: The anti-dengue IgM positivity was found to be 8.2%. The positive dengue cases were higher in male (10.5%) as compared to female (6.5%). Among different age groups, the highest positive cases (11.5 %) were from age group below 15 years followed by above 50 years age group with 8.5%. Out of 4 hospitals, the highest positive cases were in Tanahu District Hospital, Damouli (23.8%) followed by Koshi Zonal Hospital, Biratnagar (12.5%). Age and gender were found to be independent predictors. The highest numbers of dengue positive cases were in occupation group business (13.3%) followed by agriculture (11.5%). Conclusion: The dengue positivity was estimated in acute patients from different hospitals of Nepal by enzyme immunoassay and reverse transcriptase polymerase chain reaction. Therefore, the serological marker can be used to diagnose the acute patients of dengue during outbreaks. Nepal Journal of Medical Sciences | Volume 02 | Number 01 | Jan-Jun 2013 | Page 20-25 DOI: http://dx.doi.org/10.3126/njms.v2i1.7646
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Kim, Young Chan, César López-Camacho, Nallely Garcia-Larragoiti, Alan Cano-Mendez, Karina Guadalupe Hernandez-Flores, Carlos Alonso Domínguez-Alemán, Maria Antonieta Mar, Héctor Vivanco-Cid, Martha Eva Viveros-Sandoval, and Arturo Reyes-Sandoval. "Development of an E2 ELISA Methodology to Assess Chikungunya Seroprevalence in Patients from an Endemic Region of Mexico." Viruses 11, no. 5 (May 1, 2019): 407. http://dx.doi.org/10.3390/v11050407.
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Chikungunya fever is a debilitating disease caused by Chikungunya virus (CHIKV) that can result in long-lasting arthralgias. The early diagnosis of CHIKV relies on PCR during the acute infection phase to allow differential diagnosis with other co-circulating arboviruses such as dengue and Zika. Alternatively, serology can support diagnosis and provide epidemiological information on current and past outbreaks. Many commercial serological ELISA assays are based on the inactivated whole CHIKV, but their sensitivity and specificity show great variability. We produced recombinant CHIKV E2 that is suitable for ELISA assays, which was used for the serodiagnosis of CHIKV infections occurring in an arbovirus endemic Mexican region within Michoacán state. A cross-sectional study was conducted in 2016–2017; sera was obtained from 15 healthy donors and 68 patients presenting undifferentiated febrile illness. Serum samples were screened by RT-PCR and by our in-house ELISA assay. Our results indicate that IgM and IgG anti-CHIKV E2 antibodies were detected with our ELISA assay with higher sensitivity than a commercially available CHIKV ELISA kit. Our simple and sensitive ELISA assay for the serodiagnosis of CHIKV infections can be applied to population-based seroprevalence surveys and has potential for monitoring vaccine immunogenicity in CHIKV vaccine clinical trials.
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Martin, Denise A., Brad J. Biggerstaff, Becky Allen, Alison J. Johnson, Robert S. Lanciotti, and John T. Roehrig. "Use of Immunoglobulin M Cross-Reactions in Differential Diagnosis of Human Flaviviral Encephalitis Infections in the United States." Clinical and Vaccine Immunology 9, no. 3 (May 2002): 544–49. http://dx.doi.org/10.1128/cdli.9.3.544-549.2002.
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ABSTRACT To define the virus specificity of the immunoglobulin M (IgM) antibody-capture enzyme-linked immunosorbent assay (MAC-ELISA) among the medically important members of the Japanese encephalitis (JE) virus serocomplex of flaviviruses, 103 IgM-positive human serum samples from patients with confirmed West Nile (WN) virus, St. Louis encephalitis (SLE) virus, or JE virus infections were assembled and simultaneously tested against all three viral antigens in a standardized MAC-ELISA. Of the serum samples tested, 96 (93%) showed higher positive-to-negative absorbance ratios (P/Ns) with the infecting virus antigen compared to those obtained with the other two virus antigens. Of the seven specimens with higher P/Ns with heterologous virus antigens, six were from patients with SLE virus infections (the serum samples had higher levels of reactivity with WN virus antigen) and one was from a patient with a JE virus infection (this serum sample also had a higher level of reactivity with WN virus antigen). Not surprisingly, similar virus specificity was observed with WN virus-elicited IgM in cerebrospinal fluid. As shown in previous studies, a subset of these specimens was even less reactive in the MAC-ELISA with dengue virus, a member of a different flavivirus serocomplex. The degree of virus cross-reactivity did not appear to be related to days postonset, at least during the first 40 days of infection. Infections with WN virus could be correctly distinguished from infections with SLE virus on the basis of the observed anti-viral IgM cross-reactivities alone 92% of the time. Infections with SLE virus resulted in antibody that was more cross-reactive, so identification of SLE virus as the infecting agent by use of MAC-ELISA cross-reactivity alone was more problematic.
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Senaratne, T., F. Noordeen, N. Dissanayake, and KGRA Kumara. "Comparison of a rapid immunochromatography assay with an enzyme linked immunosorbent assay (ELISA) for anti-dengue virus IgM detection." Sri Lankan Journal of Infectious Diseases 4, no. 2 (October 29, 2014): 77. http://dx.doi.org/10.4038/sljid.v4i2.5925.
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Xisto, Mariana Fonseca, John Willians Oliveira Prates, Ingrid Marques Dias, Roberto Sousa Dias, Cynthia Canedo da Silva, and Sérgio Oliveira de Paula. "NS1 Recombinant Proteins Are Efficiently Produced in Pichia pastoris and Have Great Potential for Use in Diagnostic Kits for Dengue Virus Infections." Diagnostics 10, no. 6 (June 6, 2020): 379. http://dx.doi.org/10.3390/diagnostics10060379.
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Dengue is one of the major diseases causing global public health concerns. Despite technological advances in vaccine production against all its serotypes, it is estimated that the dengue virus is responsible for approximately 390 million infections per year. Laboratory diagnosis has been the key point for the correct treatment and prevention of this disease. Currently, the limiting factor in the manufacture of dengue diagnostic kits is the large-scale production of the non-structural 1 (NS1) antigen used in the capture of the antibody present in the infected patients’ serum. In this work, we demonstrate the production of the non-structural 1 protein of dengue virus (DENV) serotypes 1–4 (NS1-DENV1, NS1-DENV2, NS1-DENV3, and NS1-DENV4) in the methylotrophic yeast Pichia pastoris KM71H. Secreted recombinant protein was purified by affinity chromatography and characterized by SDS-PAGE and ELISA. The objectives of this study were achieved, and the results showed that P. pastoris is a good heterologous host and worked well in the production of NS1DENV 1–4 recombinant proteins. Easy to grow and quick to obtain, this yeast secreted ready-to-use proteins, with a final yield estimated at 2.8–4.6 milligrams per liter of culture. We reached 85–91% sensitivity and 91–93% specificity using IgM as a target, and for anti-dengue IgG, 83–87% sensitivity and 81–93% specificity were achieved. In this work, we conclude that the NS1 recombinant proteins are efficiently produced in P. pastoris and have great potential for use in diagnostic kits for dengue virus infections. The transformed yeast obtained can be used for production in industrial-scale bioreactors.
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Robinson, Makeda L., Timothy E. Sweeney, Rina Barouch-Bentov, Malaya K. Sahoo, Ana Maria Sanz, Szu-Yuan Pu, Eliana Ortiz, et al. "2565. A Novel Prognostic Gene Set for the Prediction of Severe Dengue." Open Forum Infectious Diseases 5, suppl_1 (November 2018): S72. http://dx.doi.org/10.1093/ofid/ofy209.173.
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Abstract Background There is an urgent need for the identification of biomarkers predictive of severe dengue. Single cohort transcriptomic studies have not yielded a parsimonious gene set predictive of severe dengue. We hypothesized that integration of gene expression data from heterogeneous patient populations with dengue infection would yield a set of conserved genes that is predictive of severe dengue and generalizable across cohorts. Methods Ten dengue gene expression datasets were identified in publicly available microarray repositories. A novel integrated multicohort platform was used to detect differentially expressed gene transcripts between uncomplicated and severe dengue patients and validate the identified putative signature in silico and prospectively in a new cohort of 34 dengue patients in Colombia. Dengue diagnosis was made by NS1 antigen and anti-DENV IgM antibody and confirmed by RT-PCR assays, ELISA, and IgG avidity measurements. The expression level of the signature genes was measured via microfluidic qRT-PCR assays in blood samples collected longitudinally during the course of illness. Results Using the multicohort analysis to analyze 446 peripheral blood samples of patients with dengue infection from 7 publicly available gene expression datasets, we identified a 20 gene set that predicts the development of severe dengue. We in silico validated the diagnostic power of this gene set to separate severe dengue from dengue with or without warning signs in 3 independent datasets composed of 84 samples with a global area under the ROC curve (AUC) of 0.80 [95% CI 0.68–0.88]. We prospectively validated the gene set in a new cohort composed of 34 dengue patients from Colombia with an AUC of 0.89 [95% CI 0.81–0.97]. The severity scores measured in patients with severe dengue progressively declined in longitudinal samples. Conclusion Our data indicate that the identified 20 gene signature predicts the development of severe dengue in patients prior to its onset and suggest that dengue infection itself triggers this host response. These findings may provide new insight into the pathogenesis of severe dengue and have implications for the development of a prognostic molecular assay to identify patients at risk to develop severe dengue. Disclosures T. E. Sweeney, Inflammatix, Inc.: Employee and Shareholder, Salary. P. Khatri, Inflammatix, Inc: Scientific Advisor and Shareholder, Licensing agreement or royalty.
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SA-NGASANG, A., S. ANANTAPREECHA, A. A-NUEGOONPIPAT, S. CHANAMA, S. WIBULWATTANAKIJ, K. PATTANAKUL, P. SAWANPANYALERT, and I. KURANE. "Specific IgM and IgG responses in primary and secondary dengue virus infections determined by enzyme-linked immunosorbent assay." Epidemiology and Infection 134, no. 4 (December 22, 2005): 820–25. http://dx.doi.org/10.1017/s0950268805005753.
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IgM- and IgG-capture ELISAs are widely used as diagnostic tests for confirmation of dengue virus infection. The positive rate of anti-dengue IgM and IgG detection was examined in primary and secondary dengue virus infections in the setting of a provincial hospital using IgM- and IgG-capture ELISAs. Disease day 1 was defined as the day of onset of symptoms. In total, 232 plasma samples were collected from 106 confirmed dengue cases consisting of 12 primary and 94 secondary infections. In primary infection, anti-dengue IgM was detected in 4 out of 5 samples collected on disease day 5 and in all the 21 samples collected on disease day 6 or later. Specific IgG was detected in 2 out of 5 samples collected on day 12, and in 5 out of 6 samples collected on disease days 13–15, but was not detected in samples collected on disease day 10 or earlier. In secondary infection, IgM was not detected in the samples on disease days 2 and 3, but detected in 20 out of 79 samples collected on days 4–6, in 44 out of 65 on disease days 7–11 and in 40 out of 51 samples on disease days 12–14. In contrast, specific IgG was detected in 21 out of 60 samples on disease days 4 and 5, in 13 out of 19 on disease day 6, in 62 out of 65 on disease days 7–11 and in all the samples collected on disease day 12 or later. The result indicate that seroconversion rates of IgM and IgG are different between primary and secondary infections, and suggest that detection of specific IgM and IgG is necessary for determining dengue virus infection and for differentiating primary and secondary dengue infections.
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Morales, Ivonne, Kerstin D. Rosenberger, Tereza Magalhaes, Clarice N. L. Morais, Cynthia Braga, Ernesto T. A. Marques, Guilherme Amaral Calvet, et al. "Diagnostic performance of anti-Zika virus IgM, IgAM and IgG ELISAs during co-circulation of Zika, dengue, and chikungunya viruses in Brazil and Venezuela." PLOS Neglected Tropical Diseases 15, no. 4 (April 19, 2021): e0009336. http://dx.doi.org/10.1371/journal.pntd.0009336.
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Background Serological diagnosis of Zika virus (ZIKV) infection is challenging because of the antibody cross-reactivity among flaviviruses. At the same time, the role of Nucleic Acid Testing (NAT) is limited by the low proportion of symptomatic infections and the low average viral load. Here, we compared the diagnostic performance of commercially available IgM, IgAM, and IgG ELISAs in sequential samples during the ZIKV and chikungunya (CHIKV) epidemics and co-circulation of dengue virus (DENV) in Brazil and Venezuela. Methodology/Principal findings Acute (day of illness 1–5) and follow-up (day of illness ≥ 6) blood samples were collected from nine hundred and seven symptomatic patients enrolled in a prospective multicenter study between June 2012 and August 2016. Acute samples were tested by RT-PCR for ZIKV, DENV, and CHIKV. Acute and follow-up samples were tested for IgM, IgAM, and IgG antibodies to ZIKV using commercially available ELISAs. Among follow-up samples with a RT-PCR confirmed ZIKV infection, anti-ZIKV IgAM sensitivity was 93.5% (43/46), while IgM and IgG exhibited sensitivities of 30.3% (10/33) and 72% (18/25), respectively. An additional 24% (26/109) of ZIKV infections were detected via IgAM seroconversion in ZIKV/DENV/CHIKV RT-PCR negative patients. The specificity of anti-ZIKV IgM was estimated at 93% and that of IgAM at 85%. Conclusions/Significance Our findings exemplify the challenges of the assessment of test performance for ZIKV serological tests in the real-world setting, during co-circulation of DENV, ZIKV, and CHIKV. However, we can also demonstrate that the IgAM immunoassay exhibits superior sensitivity to detect ZIKV RT-PCR confirmed infections compared to IgG and IgM immunoassays. The IgAM assay also proves to be promising for detection of anti-ZIKV seroconversions in sequential samples, both in ZIKV PCR-positive as well as PCR-negative patients, making this a candidate assay for serological monitoring of pregnant women in future ZIKV outbreaks.
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Marrero-Santos, Karla M., Manuela Beltrán, Jessica Carrión-Lebrón, Carolina Sanchez-Vegas, Davidson H. Hamer, Elizabeth D. Barnett, Luis M. Santiago, and Elizabeth A. Hunsperger. "Optimization of the Cutoff Value for a Commercial Anti-Dengue Virus IgG Immunoassay." Clinical and Vaccine Immunology 20, no. 3 (January 9, 2013): 358–62. http://dx.doi.org/10.1128/cvi.00429-12.
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ABSTRACTA commercial anti-dengue virus (anti-DENV) indirect IgG enzyme-linked immunosorbent assay (ELISA) for serological diagnosis was evaluated for its utility in determining previous DENV exposure in U.S. travelers. The Boston Area Travel Medicine Network clinics used Focus Diagnostics anti-DENV IgG ELISA to measure anti-DENV IgG antibodies in 591 pretravel specimens from U.S. residents who had traveled to countries where dengue is endemic. When using the manufacturer's index cutoff value for this ELISA, false-positive results were observed that overestimated the perceived past DENV exposure in U.S. travelers. Validation of 121 of these anti-DENV IgG results by plaque reduction neutralization test (PRNT) was used for receiver operating characteristic (ROC) curve optimization of the index cutoff value from 1 to 3.0, improving the specificity of the anti-DENV IgG ELISA from 24% to 95.7%. Additionally, previous vaccination with yellow fever virus contributed to 52.8% of the false-positive rate in the anti-DENV IgG ELISA results. Optimization of the cutoff value of the anti-DENV IgG ELISA provided better interpretation and confidence in the results and eliminated the need for confirmation by PRNT. The travel history of U.S. travelers was also useful for categorizing these travelers into groups for analysis of previous DENV exposure.
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Nascimento, Eduardo J. M., James K. George, Melissa Velasco, Matthew I. Bonaparte, Lingyi Zheng, Carlos A. DiazGranados, Ernesto T. A. Marques, and James W. Huleatt. "Development of an anti-dengue NS1 IgG ELISA to evaluate exposure to dengue virus." Journal of Virological Methods 257 (July 2018): 48–57. http://dx.doi.org/10.1016/j.jviromet.2018.03.007.
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Limothai, Umaporn, Sasipha Tachaboon, Janejira Dinhuzen, Taweewun Hunsawong, Prapapun Ong-ajchaowlerd, Butsaya Thaisomboonsuk, Stefan Fernandez, et al. "Dengue pre-vaccination screening test evaluation for the use of dengue vaccine in an endemic area." PLOS ONE 16, no. 9 (September 10, 2021): e0257182. http://dx.doi.org/10.1371/journal.pone.0257182.
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Background The dengue vaccine (Dengvaxia) is only recommended for individuals with prior dengue infection (PDI). This study aimed to perform a serosurvey to inform decision-making for vaccine introduction and identify appropriate target populations. We also evaluated the performance of the serological tests using plaque reduction neutralization test (PRNT) as a reference test in identifying PDI to determine suitability for pre-vaccination screening. Methods We enrolled 115 healthy individuals between 10 and 22 years of age living in the Ratchaburi province of Thailand. The serum samples were tested by PRNT to measure the prevalence and concentration of serotype-specific neutralizing antibodies. The performance of the IgG rapid diagnostic test (RDT, SD Bioline, Korea) and IgG enzyme-linked immunosorbent assay (ELISA, EUROIMMUN, Germany) in identifying PDI were evaluated by using PRNT as a reference method. Results Ninety-four (81.7%) individuals neutralized one or more dengue serotypes at a titer threshold greater than or equal to 10. Multitypic profiles were observed in 70.4% of the samples which increased to 91.9% in subjects aged 19–22. Among monotypic samples, the highest proportion was reactive against DENV-1 followed by DENV-2, DENV-3, and DENV-4. The highest anti-dengue antibody titers were recorded against DENV-1 and increased with age to a geometric mean NT50 titer (GMT) of 188.6 in the 19–22 age group. While both RDT and ELISA exhibited 100% specificity, RDT demonstrated low sensitivity (35%) with ELISA displaying much greater sensitivity (87%). Conclusions Almost 80% of adolescents and youth in Ratchaburi province had already been exposed to one or more of the dengue virus serotypes. The dengue IgG RDT displayed low sensitivity and is likely not be suitable for dengue pre-vaccination screening. These results support the use of IgG ELISA test for dengue vaccination in endemic areas.
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Mahmood, Shahid, Hiba Nabeel, Saadia Hafeez, Urooj Zahra, and Hammad Nazeer. "Seroprevalence of Dengue IgG Antibodies among Healthy Adult Population in Lahore, Pakistan." ISRN Tropical Medicine 2013 (September 12, 2013): 1–6. http://dx.doi.org/10.1155/2013/521396.
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Background. Dengue is a mosquito-borne flavivirus infection prevalent in tropical and subtropical regions around the world. Aim of this study was to determine seroprevalence of anti-dengue IgG antibodies in healthy adult population of Lahore and also describe risk factors in relation to dengue seropositivity. Methods. In this cross-sectional study, 274 healthy adult individuals aged 15 years and above were randomly selected using multistage sampling technique. These individuals were interviewed between July–September 2012, using a semistructured questionnaire, followed by drawing 3 mL of their venous blood for dengue IgG test. Nova Tech ELISA kit with sensitivity and specificity of 96.5% and 97.5%, respectively, was used for serology. Results. Out of 274 participants, 184 (67.2%) were found to be positive for dengue IgG antibodies. Seroprevalence was higher among individuals with poor awareness about potential breeding sites for dengue mosquito (63.6%), followed by the subjects who had poor knowledge about dengue signs/symptoms and complications (52.2% and 68.5%, resp.). Conclusion. About two-third of healthy population of Lahore was also seropositive for anti-dengue IgG during July–September 2012, indicating a considerable burden of subclinical dengue infection in the city. Males were predominantly affected than the females. We found no statistical association between dengue IgG seropositivity and socioeconomic status, occupation, and knowledge about the disease.
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Weissbach, Fabian H., and Hans H. Hirsch. "Comparison of Two Commercial Tick-Borne Encephalitis Virus IgG Enzyme-Linked Immunosorbent Assays." Clinical and Vaccine Immunology 22, no. 7 (April 29, 2015): 754–60. http://dx.doi.org/10.1128/cvi.00096-15.
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ABSTRACTDespite the availability of protective vaccines, tick-borne encephalitis virus (TBEV) infections have been increasingly reported to the European Centre for Disease Prevention and Control in the past 2 decades. Since the diagnosis of TBEV exposure relies on serological testing, we compared two commercial enzyme-linked immunosorbent assays (ELISAs), i.e., Immunozym FSME IgG assay (ELISA-1) and Euroimmun FSME Vienna IgG assay (ELISA-2). Both assays use whole TBEV antigens, but they differ in viral strains (Neudoerfl for ELISA-1 and K23 for ELISA-2) and cutoff values. In testing of samples from 398 healthy blood donors, ELISA-1 showed higher reactivity levels than ELISA-2 (P< 0.001), suggesting different assay properties. This finding was supported by Bland-Altman analysis of the optical density at 450 nm (OD450) (mean bias, +0.32 [95% limits of agreement, −0.31 to +0.95]) and persisted after transformation into Vienna units. Concordant results were observed for 276 sera (69%) (44 positive and 232 negative results). Discordant results were observed for 122 sera (31%); 15 were fully discordant, all being ELISA-1 positive and ELISA-2 negative, and 107 were partially discordant (101 being ELISA-1 indeterminate and ELISA-2 negative and 6 having positive or indeterminate reactivity in both ELISAs). Neutralization testing at a 1:10 dilution yielded positive results for 33 of 44 concordant positive sera, 1 of 15 fully discordant sera, and 1 of 33 partially discordant sera. Indirect immunofluorescence testing revealed high antibody titers of ≥100 for yellow fever virus in 18 cases and for dengue virus in one case, suggesting that cross-reactivity contributed to the ELISA-1 results. We conclude that (i) cross-reactivity among flaviviruses remains a limitation of TBEV serological testing, (ii) ELISA-2 revealed reasonable sensitivity and specificity for anti-TBEV IgG population screening of human sera, and (iii) neutralization testing is most specific and should be reserved for selective questions.
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Libraty, Daniel H., Lei Zhang, AnaMae Obcena, Job D. Brion, and Rosario Z. Capeding. "Anti-dengue virus envelope protein domain III IgG ELISA among infants with primary dengue virus infections." Acta Tropica 142 (February 2015): 103–7. http://dx.doi.org/10.1016/j.actatropica.2014.11.009.
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García Rojas, Claudia Lorena, Dalgys Martínez, Dolly Castro, Doris Martha Salgado, Carlos Narváez, Leonardo Puerta, and Jairo Antonio Rodríguez. "Anticuerpos IgG4 específicos anti Aedes aegypti como factor protector en niños con dengue grave." Revista Med 27, no. 2 (July 1, 2020): 11–20. http://dx.doi.org/10.18359/rmed.3548.
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Introducción: la infección por el virus del dengue es un problema de salud pública mundial. El virus es transmitido por la picadura de mosquitos del género Aedes. Las proteínas de la saliva del vector Aedes aegypti inducen anticuerpos IgE e IgG4 específicos, cuya relación con la gravedad del dengue aún es desconocida. Objetivo: evaluar la asociación entre anticuerpos IgE e IgG4 específicos anti A. aegypti con la gravedad de la infección por dengue. Método: se realizó un estudio transversal en el que se incluyeron 16 niños con dengue grave (dg), 15 niños con dengue con signos de alarma (dcsa) y 26 niños sanos, todos menores de 15 años. Se determinaron niveles séricos de IgE e IgG4 específicas de A. aegypti; también se cuantificó vegf, sst2 y vegfr1 por elisa. Para las variables cualitativas se calcularon proporciones y odds ratio (or); en las variables cuantitativas se hallaron medianas, rango intercuartílico y se utilizó la prueba U Mann Whitney. Resultados: la oportunidad de los niños de tener dg con niveles séricos de IgG4 específica mayores de 0,5 od es 78 % menor [or=0,22] (ic de 95 % de 0,06-0,77), comparado con la oportunidad de tener dg con niveles séricos de IgG4 específica menores de 0,5 od. Plaquetas (p=0,0002) y vefg (p=0,003) más elevado en los pacientes con dcsa y sst2 fue más alto en el dg (p=0,004). Conclusión: niveles de anticuerpos de IgG4 anti A. aegypti se relacionan con menor gravedad clínica del dengue.
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Tan, L. K., C. L. Too, A. F. Nurul-Aain, A. A. Siti-Aisyah, S. Wahinuddin, A. Osman, I. S. Lau, et al. "OP0096 EXPOSURE TO DENGUE INFECTION DO NOT RAISE RISK OF RHEUMATOID ARTHRITIS: FINDINGS FROM THE MALAYSIAN EPIDEMIOLOGICAL INVESTIGATION OF RHEUMATOID ARTHRITIS (MYEIRA) CASE-CONTROL STUDY." Annals of the Rheumatic Diseases 80, Suppl 1 (May 19, 2021): 53.1–53. http://dx.doi.org/10.1136/annrheumdis-2021-eular.1684.
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Background:Dengue infection is associated with joints pain mimicking disease onset symptom of rheumatoid arthritis (RA). However, there is lack of epidemiological studies on exposure to dengue infection and risk of future RA.Objectives:We investigated the relationship between exposure to dengue infection and risk of developing different subsets of RA, defined by the presence of anti-citrullinated peptide antibody (ACPA) in the multi-ethnic Malaysian population.Methods:Serum samples from 1,235 RA cases (i.e. 516 Malay, 254 Chinese, 405 Indians and 60 others/mixed-ethnicity) and 1,624 epidemiological matched population-based controls (i.e. 1,023 Malay, 208 Chinese, 297 Indians and 96 others/mixed-ethnicity) were assayed for presence of dengue IgG antibody using World Health Organization recommended ELISA kits. Positive results of dengue IgG antibodies indicates previous exposure to dengue infection(s). We performed chi-square and Mann-Whitney U analysis to determine the association of ever-exposed dengue infection with ACPA-positive/ACPA-negative RA and to investigate the antibody frequency and levels among the studied populations.Results:We observed high occurrence of dengue IgG antibody in the overall RA cases (79.7%) and matched controls (77.3%), with no significant differences detected between the ACPA subsets of RA. Ethnicity stratification analysis revealed a decrease risk of developing ACPA-positive RA in the Indian patients with positive dengue IgG antibody (OR=0.59, 95% CI=0.37-0.94, p=0.03), and in particular patients with elevated level of dengue IgG antibody (OR=0.44, 95% CI=0.25-0.78, p<0.05). On the other hand, the significant decrease mean levels of dengue IgG antibody were observed in the ACPA-positive RA subset for all three major ethnic groups (i.e. Malay, p<0.0001, Chinese, p<0.01 and Indian<0.05) (Figure 1). No association was observed between presence of dengue IgG antibody and ACPA-negative RA subset.Figure 1.Comparison of mean dengue IgG antibody level between ever-exposed dengue infection RA cases, stratified by ACPA status. Comparison of median dengue IgG antibody level between the ever-exposed dengue infection ACPA-positive RA and normal controls in the four ethnic groups. The red line indicates the mean level of dengue IgG antibody levelConclusion:Our findings demonstrated that exposure to dengue infection do not increase the risk of developing future RA in the multi-ethnic Malaysian population. The inverse associations observed in the Indian ethnic group are in line with the other studies investigating exposure to viral infection and risk of RA.References:[1]Sherina et al (2017) Low levels of antibodies against common viruses associate with anti-citrullinated protein antibody-positive rheumatoid arthritis; implications for disease aetiology. Arthritis Research & Therapy 2017, 19:2169[2]Gissel García et. al. (2011) Long-term persistence of clinical symptoms in dengue-infected persons and its association with immunological disorders. International Journal of Infectious Diseases 15 (2011) e38–e43Acknowledgements:The authors would like to thank the Director General of Health, Ministry of Health Malaysia for supporting this study. The authors are also indebted to participants for their kind participation. This study was financially supported by the Ministry of Health, Malaysia (JPP-IMR 17-025) and the short-term research grant by UniKL RCMP (str16037).Disclosure of Interests:None declared