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Статті в журналах з теми "RT-PCR kit"
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Lemasova, L. V., G. A. Tkachenko, E. V. Prokhvatilova, L. I. Belitskaya, D. V. Viktorov, and A. V. Toporkov. "ASSESSMENT OF THE POSSIBILITY OF APPLICATION IN LABORATORY PRACTICE OF REAGENT KIT FOR DIAGNOSIS OF GLANDERS AND MELIOIDOSIS BY REAL-TIME POLYMERASE CHAIN REACTION." Russian Clinical Laboratory Diagnostics 64, no. 11 (November 15, 2019): 700–704. http://dx.doi.org/10.18821/0869-2084-2019-64-11-700-704.
Повний текст джерелаАнотація:
The reagent kit AmpligenBurk-mallei/pseudomallei-RT PCR is designed for detecting in vitro diagnostics and differentiate the DNA of glanders and melioidosis pathogens by real-time multiplex PCR in biological (clinical) material and cultures of microorganisms, as well as environmental objects and solid food products (rice). During clinical testing diagnostic value of reagent kit AmpligenBurk-mallei/pseudomallei-RT PCR has been studied. Based on the results obtained, a high analytical sensitivity (1×103 microbe cells/ml) and specificity (100%) of PCR-RT with the developed reagent kit were established, regardless of the type of material being studied. The diagnostic sensitivity of PCR-RT using a set of reagents was at least 98.0% and specificity at least 99%. The stages of state examination have been completed, a registration certificate has been obtained at Roszdravnadzor, production, sale and use of reagent kit in medical laboratory practice have been permitted.
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Goncharova, E. V., A. E. Donnikov, V. V. Kadochnikova, S. A. Morozova, M. N. Boldyreva, I. S. Galkina, and D. V. Blinov. "Real-time RT-PCR diagnostics of virus causing COVID-19." FARMAKOEKONOMIKA. Modern Pharmacoeconomic and Pharmacoepidemiology 13, no. 1 (April 24, 2020): 52–63. http://dx.doi.org/10.17749/2070-4909.2020.13.1.52-63.
Повний текст джерелаАнотація:
Aim: the study was aimed to develop a reagent kit for the real-time RT-PCR diagnostics of virus causing COVID-19.Materials and Methods. Three target sites were chosen in the genome SARS-CoV-2. The testing included 220 samples, 48 artificially created positive samples (made from patients’ biomaterial) and 172 clinical samples (scrapes from nasal and pharyngeal cavities, bronchoalveolar lavage, expectoration, endotracheal/nasopharyngeal aspirate, feces, post-mortem material), obtained from two medical centers. Preliminary, the obtained biomaterial was analyzed with a reagent kit of comparison. The evaluation was performed with a confidential interval CI 95%. The calculation of CI for the sensitivity and specificity was made based on the distribution of χ2.Results. The authors developed a technology of novel coronavirus infection (COVID-19) real-time RT-PCR diagnostics for the application in practical healthcare and proposed the variants of testing at all the stages (preanalytical, analytical, and post-analytical, including automated results processing). The proposed reagent kit meets the requirements of the World Health Organization and the Ministry of Healthcare of the Russian Federation. The study results demonstrated high sensitivity and specificity. The sensitivity was 100% (95% CI) 95.6–100%; the specificity was 100% (95% CI) 96.7–100%.Conclusion. The proposed reagent kit was registered in the RF as a medical product; the registration certificate No. RZN 2020/9948 dated 01.04.2020. The application of the reagent kit in network laboratories will provide patients with access to testing for the virus causing COVID-19 and contribute to quick differential diagnostics, improvement of pandemic control, and accurate statistics on the spread of the virus.
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Hecht, Leonie-Sophie, Angeles Jurado-Jimenez, Markus Hess, Hussein El Halas, Gregor Bochenek, Hala Mohammed, Fahd Alzahrani, et al. "Verification and diagnostic evaluation of the RealStar® Middle East respiratory syndrome coronavirus (N gene) reverse transcription-PCR kit 1.0." Future Microbiology 14, no. 11 (July 2019): 941–48. http://dx.doi.org/10.2217/fmb-2019-0067.
Повний текст джерелаАнотація:
Aim: We report the diagnostic evaluation of a confirmatory reverse transcription-PCR (RT-PCR) kit targeting the Middle East respiratory syndrome coronavirus (MERS-CoV) N gene. Material & methods: 33 patient samples from two collections sites in Riyadh, Saudi Arabia, which were pre-characterized via real-time RT-PCR targeting MERS-CoV orf1a and upE, and were tested using the MERS-CoV N gene, as a confirmatory assay. This diagnostic procedure follows a two-step diagnostics scheme, recommended by the WHO. Results: 18/33 samples tested positive, 11/33 tested negative for MERS-CoV RNA and 2/33 showed uncertain results. Conclusion: The results suggest, that the RealStar® MERS-CoV (N gene) RT-PCR kit 1.0 can be considered a suitable and reliable confirmatory assay in combination with the RealStar MERS-CoV RT-PCR kit 1.0 according to the diagnostic scheme recommended by WHO.
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Hu, ZB, W. Ma, CC Uphoff, H. Quentmeier, and HG Drexler. "c-kit expression in human megakaryoblastic leukemia cell lines." Blood 83, no. 8 (April 15, 1994): 2133–44. http://dx.doi.org/10.1182/blood.v83.8.2133.2133.
Повний текст джерелаАнотація:
Abstract A panel of 164 continuous human leukemia-lymphoma cell lines was analyzed for expression of c-kit using Northern blotting and reverse transcriptase-polymerase chain reaction (RT-PCR). The c-kit transcripts were detectable in cell lines assigned to the myeloid (in 7 of 29 by Northern blotting and in 4 of 8 by RT-PCR), monocytic (in 1 of 24 by Northern blotting and in 3 of 6 by RT-PCR), erythroid (in 6 of 8 by Northern blotting and in 5 of 5 by RT-PCR), and megakaryoblastic (in 10 of 10 by Northern blotting) lineages, c-kit expression was not seen by Northern blotting or RT-PCR analysis in any of the 93 lymphoid leukemia, myeloma, or lymphoma cell lines. Treatment of four megakaryoblastic cell lines with protein kinase C activators (phorbol ester 12-O-tetradecanoylphorbol 13-acetate and Bryostatin 1) led to terminal differentiation as assessed by morphologic alterations, changes in the surface marker profile, and growth arrest. These effects were associated with enhanced c-kit mRNA expression. Exposure to all- trans retinoic acid down-regulated c-kit mRNA levels, while simultaneously causing morphologic alterations in all four cell lines. Stimulation with growth factors (interleukin-3, granulocyte macrophage- colony stimulating factor, and insulin-like growth factors I and II), used to assess any role of c-kit in proliferative processes, did not lead to significant upregulation or downregulation of c-kit expression. The finding of constitutive and high expression of c-kit mRNA in all megakaryoblastic leukemia cell lines and its modulation by various reagents might further contribute to the understanding of megakaryopoietic proliferation, differentiation, and leukemogenesis.
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Hu, ZB, W. Ma, CC Uphoff, H. Quentmeier, and HG Drexler. "c-kit expression in human megakaryoblastic leukemia cell lines." Blood 83, no. 8 (April 15, 1994): 2133–44. http://dx.doi.org/10.1182/blood.v83.8.2133.bloodjournal8382133.
Повний текст джерелаАнотація:
A panel of 164 continuous human leukemia-lymphoma cell lines was analyzed for expression of c-kit using Northern blotting and reverse transcriptase-polymerase chain reaction (RT-PCR). The c-kit transcripts were detectable in cell lines assigned to the myeloid (in 7 of 29 by Northern blotting and in 4 of 8 by RT-PCR), monocytic (in 1 of 24 by Northern blotting and in 3 of 6 by RT-PCR), erythroid (in 6 of 8 by Northern blotting and in 5 of 5 by RT-PCR), and megakaryoblastic (in 10 of 10 by Northern blotting) lineages, c-kit expression was not seen by Northern blotting or RT-PCR analysis in any of the 93 lymphoid leukemia, myeloma, or lymphoma cell lines. Treatment of four megakaryoblastic cell lines with protein kinase C activators (phorbol ester 12-O-tetradecanoylphorbol 13-acetate and Bryostatin 1) led to terminal differentiation as assessed by morphologic alterations, changes in the surface marker profile, and growth arrest. These effects were associated with enhanced c-kit mRNA expression. Exposure to all- trans retinoic acid down-regulated c-kit mRNA levels, while simultaneously causing morphologic alterations in all four cell lines. Stimulation with growth factors (interleukin-3, granulocyte macrophage- colony stimulating factor, and insulin-like growth factors I and II), used to assess any role of c-kit in proliferative processes, did not lead to significant upregulation or downregulation of c-kit expression. The finding of constitutive and high expression of c-kit mRNA in all megakaryoblastic leukemia cell lines and its modulation by various reagents might further contribute to the understanding of megakaryopoietic proliferation, differentiation, and leukemogenesis.
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Deng, Ming Y., He Wang, Gordon B. Ward, Tammy R. Beckham, and Thomas S. McKenna. "Comparison of Six RNA Extraction Methods for the Detection of Classical Swine Fever Virus by Real-Time and Conventional Reverse Transcription–PCR." Journal of Veterinary Diagnostic Investigation 17, no. 6 (November 2005): 574–78. http://dx.doi.org/10.1177/104063870501700609.
Повний текст джерелаАнотація:
Six RNA extraction methods, i.e., RNAqueous kit, Micro-to-midi total RNA purification system, NucleoSpin RNA II, GenElute mammalian total RNA kit, RNeasy mini kit, and TRIzol LS reagent, were evaluated on blood and 7 tissues from pig infected with classical swine fever virus (CSFV). Each of the 6 extraction methods yielded sufficient RNA for positive results in a real-time reverse transcription–PCR (RT-PCR) for CSFV, and all RNA, except the one extracted from blood by TRIzol LS reagent, yielded positive results in both a conventional RT-PCR for CSFV and a conventional RT-PCR for an endogenous gene encoding β-actin. The RNA extracted from blood by TRIzol LS reagent became positive in both conventional RT-PCR assays when it was diluted to 1:2, 1:4, or up to 1:64 in nuclease-free water. It is concluded that all 6 methods are more or less useful for the detection of CSFV by real-time and conventional RT-PCR in swine blood and tissues. However, some of the 6 reagents offer certain advantages not common to all 6 extraction procedures. For example, RNA extracted by the TRIzol LS reagent constantly had the highest yield; that by the RNAqueous kit had the highest A260/A280 ratio for almost all samples; and that by the NucleoSpin RNA II and the GenElute mammalian total RNA kit was most likely to be free of contaminations with genomic DNA.
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Ningrum, Esti Prasetya, Sedyo Hartono, Sri Sulandari, and Susamto Somowiyarjo. "Multiplex RT-PCR Assay for Crinivirus Detection Using RNA Prepared from Three Extraction Methods on Tomato Plant." Jurnal Perlindungan Tanaman Indonesia 23, no. 2 (December 3, 2019): 250. http://dx.doi.org/10.22146/jpti.36558.
Повний текст джерелаАнотація:
Tomato infectious chlorosis virus (TICV) and Tomato chlorosis virus (ToCV) are members of the Crinivirus genus that induces yellowing symptoms in tomato plants. Detection of both viruses is generally carried out singly, thus it is necessary to develop a fast, accurate and efficient detection method to detect multiple viruses simultaneously in an effort to determine the suitable disease management strategies. This study was aimed to detect both viruses using the multiplex RT-PCR method and evaluate three methods of total RNA preparation used from tomato plants as RT-PCR templates. The methods evaluated were simple direct tube (SDT), simple dsRNA, and commercial kit (RNeasy Plant Mini Kit) as a comparison. The total source of RNA came from Crinivirus symptomatic tomato leaves from Kopeng, and Ketep (Central Java); Pakem (Yogyakarta); Malang (East Java); and Bogor (West Java). Single RT-PCR and multiplex RT-PCR using specific primers CPd I/CPd II and ToCV CF/ToCV CR with DNA band targets of 760 bp and 360 bp. The SDT and dsRNA methods have been successful in obtaining total RNA and viral RNA from tomato leaf samples. Total RNA RT-PCR with simple SDT and dsRNA methods followed by multiplex RT-PCR produces specific DNA band intensities that are comparable to Kit. RNA preparation with SDT and simple dsRNA methods is a simple, fast, easy and affordable method in providing templates for RT-PCR. Multiplex RT-PCR technique using two pairs of specific primers CPd I/CPd II and ToCV CF/ToCV CR is suitable for simultaneous testing of TICV and ToCV.
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Demeke, Tigst, Jemima Malabanan, Michelle Holigroski, and Monika Eng. "Effect of Source of DNA on the Quantitative Analysis of Genetically Engineered Traits Using Digital PCR and Real-Time PCR." Journal of AOAC INTERNATIONAL 100, no. 2 (March 1, 2017): 492–98. http://dx.doi.org/10.5740/jaoacint.16-0284.
Повний текст джерелаАнотація:
Abstract Seven commercially available DNA extraction kits were compared with a cetyltrimethylammonium bromide (CTAB)method to determine the suitability of the extracted DNA for RainDrop digital PCR (dPCR) and real-time PCR (RT-PCR) quantification of OXY235 canola, FP967 flax, and DP305423 soybean (spiked at the 0.1% level). For the kits, the highest amount of DNA extracted from a 0.2 g sample was obtained using OmniPrep for Plant for flax andDNeasy mericon Food for canola and soybean. For canola, DNA extracted with the Fast ID Genomic DNA Extraction Kit, FastDNA Spin Kit, GM Quicker 2, NucleoSpin Food, and DNeasy mericon Food was suitable for dPCR and RT-PCR. For flax, DNA extracted with Fast ID, FastDNA Spin Kit, OmniPrep for Plant, and NucleoSpin Food was suitable for RT-PCR. However, only Fast ID yielded DNA suitable for dPCR. For soybean, DNA extracted with five and six of the seven DNA extraction kits was suitable for dPCR and RT-PCR, respectively. Overall, Fast ID provided reliable results regardless of species or analysis method used. Canola, flax, and soybean DNA extracted with the CTAB method and then purified were suitable for both dPCR and RT-PCR. This is the first report showing the effect of different DNA extraction methods on the absolute quantification of genetically engineered traits using dPCR.
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Otter, Jonathan A., Nancy L. Havill, and John M. Boyce. "Evaluation of Real-Time Polymerase Chain Reaction for the Detection of Methicillin-ResistantStaphylococcus aureuson Environmental Surfaces." Infection Control & Hospital Epidemiology 28, no. 8 (August 2007): 1003–5. http://dx.doi.org/10.1086/519207.
Повний текст джерелаАнотація:
We compared real-time polymerase chain reaction (RT-PCR) with in vitro culture for detecting methicillin-resistantStaphylococcus aureusin samples from environmental surfaces. The sensitivity of RT-PCR, compared with culture, was 92.5%, and the specificity was 51.4%. Because of poor specificity, the RT-PCR kit tested is not suitable for the detection of MRSA on hospital surfaces.
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Stephenson, Patrick, David Crabtree, Katharine Evans, Heikki Salavirta, Pius Brzoska, Ana-Maria Leonte, Amanda Manolis, and Daniele Sohier. "Validation of the Thermo Scientific™ SARS-CoV-2 RT-PCR Detection Workflow for the Detection of SARS-CoV-2 from Stainless-Steel Environmental Surface Swabs: AOAC Performance Tested MethodSM 012103." Journal of AOAC INTERNATIONAL 104, no. 4 (April 5, 2021): 935–47. http://dx.doi.org/10.1093/jaoacint/qsab050.
Повний текст джерелаАнотація:
Abstract Background The Thermo Scientific™ SARS-CoV-2 reverse transcription–polymerase chain reaction (RT-PCR) Detection Workflow, packaged with Applied Biosystems™ TaqMan™ 2019-nCoV Assay Kit v1 targets three different SARS-CoV-2 genomic regions in a single RT-PCR reaction. Objective To validate the Thermo Scientific SARS-CoV-2 RT-PCR Workflow, for the detection of SARS-CoV-2 virus on stainless-steel surfaces as part of the AOAC Performance Tested MethodSM Emergency Response Validation program. Method The Applied Biosystems TaqMan 2019-nCoV Assay Kit v1, as part of the Thermo Scientific SARS-CoV-2 RT-PCR Workflow, was evaluated for specificity using in silico analysis of 15 764 SARS-CoV-2 sequences and 65 exclusivity organisms. The Thermo Scientific SARS-CoV-2 RT-PCR Workflow was evaluated in an unpaired study for one environmental surface (stainless steel) and compared to the U.S. Centers for Disease Control and Prevention 2019-Novel Coronavirus RT-PCR Diagnostic Panel, Instructions for Use (Revision 4, Effective 6/12/2020). Results In silico analysis showed that, of the 15 756 target SARS-CoV-2 genomes analyzed, 99% of the strains/isolates are perfectly matched to at least two of the three assays, and more than 90% have 100% homology to all three assays (ORF1ab, N-gene, S-gene) in the SARS-CoV-2 Kit. None of the 65 non-target strain genomes analyzed showed matching sequences. In the matrix study, the Thermo Scientific SARS-CoV-2 workflow showed comparable detection to the centers of disease control and prevention (CDC) method. Conclusions The Thermo Scientific SARS-CoV-2 RT-PCR Workflow is an effective procedure for detection of RNA from SARS-CoV-2 virus from stainless steel. Highlights The workflow provides equivalent performance results with the two tested RNA extraction platforms and the two tested RT-PCR instruments.
Дисертації з теми "RT-PCR kit"
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Milhem, Clara. "Caractérisation et validation d'une signature moléculaire des lymphocytes T régulateurs humains et de leur microenvironnement : mise en place d'un test compagnon." Thesis, Université de Lille (2022-....), 2022. http://www.theses.fr/2022ULILS008.
Повний текст джерелаАнотація:
Les lymphocytes T régulateurs (Treg) sont une sous-population de cellules immunitaires connues pour leur rôle prépondérant dans la modulation de la réponse immunitaire innée et adaptative. Ils ont un rôle majeur dans la physiologie de l’organisme, mais également dans la mise en place des pathologies comme les inflammations chroniques, les maladies auto-immunes et les cancers. L’intérêt de mieux caractériser et suivre l’évolution des Tregs n’est plus à démontrer, qui plus est à l’ère des immunothérapies. En ce sens, notre équipe a établi une signature moléculaire unique des Tregs, basée sur l’expression de 81 gènes permettant de les caractériser et d’évaluer leur modulation. Ainsi, la validation de la signature moléculaire aboutira à l’établissement d’un kit compagnon couplé à un algorithme de lecture. Cela permettra de prévenir de façon fiable et rapide les effets indésirables de nouvelles molécules, de la recherche jusqu'à la clinique, vis-à-vis du système immunitaire.La première partie des résultats de qPCR sur les Tregs isolés d’individus sains a permis de conclure à la stabilité d’expression pour 84% des gènes de la signature. Ce ratio monte jusqu’à 92% pour l’évaluation de la stabilité d’expression des gènes dans les PBMC des mêmes donneurs. Ces résultats, très prometteurs, nécessitent d’être confirmés sur une plus grande population. Ainsi, l’étude d’une corrélation mathématique entre les niveaux d’expressions géniques des Tregs isolés et des PBMC autologues a été entrepris. Cette étude a permis de montrer que l’expression de 11 gènes est corrélée de manière linéaire à plus de 70% entre Tregs isolés et PBMC autologues.Dans un second temps, ce travail a permis d’évaluer la réponse de la signature face à des agents modulateurs décrits pour activer ou inhiber les Tregs. Cette étape a validé la bonne réactivité de certains gènes face aux agents modulateurs. C’est le cas de 4 gènes, qui diminuent significativement quand l’activité suppressive des Tregs est inhibée par un anticorps monoclonal anti-galectine-9. Néanmoins, l’analyse de variance multivariée n’a pas permis de montrer que les variations d’expression observées sont exclusivement dues au traitement sur la base des 10 donneurs, il semble que la variabilité interindividuelle entre en jeu dans l’analyse de certains groupes de gènes.Enfin, le comportement de la signature a été évalué chez 4 patients, inclus dans un essai clinique pour le traitement de différents types de cancer par radiothérapie stéréotaxique hypo fractionnée à haute dose. Encore une fois, les résultats obtenus sont très prometteurs puisque l’évolution biologique et clinique des patients est corrélée avec l’évolution d’expression des gènes de la signature. Ces données constituent une première confirmation de l’utilité d’une telle signature dans l’immuno-monitoring des patients.Ces études sont très encourageantes pour le développement d’un algorithme de lecture automatisé des résultats. Ainsi, le kit pourra être positionnable sur le marché de la recherche fondamentale jusqu’en clinique, pour l’évaluation des Tregs et leurs microenvironnements dans tous types d’étudesRegulatory T cells (Treg) are an immune cell subpopulation known for their important role in innate and adaptive immune response contraction. They have a major role in physiology, but also in pathologies outcome such as chronic inflammations, autoimmune diseases and cancers. A better characterization and follow of Treg evolution are clearly needed, especially in the era of immunotherapies. Our team has established a unique Treg molecular signature based on the expression of 81 genes, allowing to characterize and to evaluate Treg modulation. Thus, the molecular signature validation will lead to the establishment of a companion kit coupled with a reading algorithm. This will allow to reliably and rapidly prevent the adverse effects of new molecules, from research to the clinic, regarding the immune system.The first part of the RT-PCR results on healthy donors isolated Tregs allowed to conclude that 84% of the genes have stable expression. This ratio rises to 92% for the stability evaluation of gene expression in PBMC from the same donors. These results are very promising, but need to be confirmed on a larger population. To go further the presence of a mathematical correlation between isolated Tregs and autologous PBMC gene expression levels, have been researched. This study showed that the expression of 11 gene are linearly correlated beyond 70%. Afterward, the signature response to modulating agents described to activate or inhibit Tregs have been asses. Here, the genes good response to modulating agents is proved: 4 gene expression decrease significantly when Tregs suppressive activity is inhibited by an anti-galectin-9 monoclonal antibody. Nevertheless, multivariate analysis of variance failed to show that the expression variations are exclusively due to treatment with 10 donors, it seems that interindividual variability has a role in some gene response to modulator. Finally, the signature behavior was evaluated in 4 patients, included in a clinical trial for cancer treatment by high dose hypo fractionated stereotactic radiotherapy. Once again, the results obtained are very promising since the biological and clinical patients’ evolution is correlated with the gene expression modulation. These data constitute a first confirmation of the usefulness of such a signature in the immuno-monitoring of patients.These outcomes are very encouraging for the development of an automated reading algorithm. Thus, the kit could be positioned on the market from basic research to the clinic, for the evaluation of Tregs and their micro environments in all types of studies
Частини книг з теми "RT-PCR kit"
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Pai, Girish, Mansee Thakur, Harapriya Kar, and D. S. Joshi. "Validation of in House PCR Using IS6110 for Detection of M. tuberculosis and Its Comparison with ZN Staining, Cultures and RT PCR Kit Methods." In Techno-Societal 2016, 599–609. Cham: Springer International Publishing, 2017. http://dx.doi.org/10.1007/978-3-319-53556-2_61.
Повний текст джерела
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Sunil Patankar, Rutuja, and Vasudeo Pandharinath Zambare. "Development of RT-PCR Based Diagnosis of SARS-CoV-2." In Biotechnology to Combat COVID-19 [Working Title]. IntechOpen, 2021. http://dx.doi.org/10.5772/intechopen.96823.
Повний текст джерелаАнотація:
In the 2020, COVID-19 pandemic disease created an havoc situation world widely and mainly caused by Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2). It has been challenging task for researchers, scientists and medico-pharmaceutical organisations to find out rapid and reliable diagnosis methods. Among the all testing services, a Reverse Transcription Polymerase Chain Reaction (RT-PCR) is the more accurate, rapid and authenticated molecular technique used for most of the diagnosis of major diseases. It has been a global priority to fix the rapid diagnosis method to combat against the pandemic COVID-19. Thus, the present chapter mainly focussing on the progress of RT-PCR method development though various processes of data collection on isolation of whole genome sequence, its primer and method designing. In this scenario, India suddenly become the global leader for vaccine development and hence the challenges and RT-PCR kit development in India and rest of the world has been be discussed. World wide many Government and private agencies and industries have taken an initiative for diagnosis of SARS-CoV-2 hence this chapter also summarised the scope of RT-PCR to combat pandemic situation in future.
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Swetha.M and Rajendiran. M. "Effective Early Stage Detection of COVID-19 Using Deep Learning." In Advances in Parallel Computing. IOS Press, 2021. http://dx.doi.org/10.3233/apc210037.
Повний текст джерелаАнотація:
The Corona virus Disease 2019 (COVID-19), which was formerly called as 2019 Novel Corona Virus[1] is a breath taking disease. It had its impact on millions of lives across the world. At present, as of March 2021, the rate of infection has declined throughout different parts of the world [2]. But it has been warned by scientists that this deadly disease can have its second wave over a period of time. Also, there is a possibility of this covid-19 to become a seasonal disease [3]. In such case, premature diagnosis of this virus is essential in order to save many lives. A kit called RT-PCR has been employed to detect the presence of this virus [4]. However, this method of prognosis takes time depending on the locality of the infected person [5]. This leads to the proliferation of the infection. Hence, an alternate procedure should be unearthed, which diagnose this disease within a short span of time. In this paper, a Deep Learning concept has been proposed which aids the timely detection of the corona virus infection. This, inturn reduces the spreading rate of the infection and decreases mortality rate.
Тези доповідей конференцій з теми "RT-PCR kit"
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Fleming, Paul, and Tara Dalton. "One-Step Reverse-Transcription PCR on a High-Throughput Micro-Fluidic Device." In ASME 2009 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2009. http://dx.doi.org/10.1115/sbc2009-206623.
Повний текст джерелаАнотація:
One step reverse-transcription polymerase chain reaction (RT-PCR) assays are an attractive option for further automating gene detection assays. One-step assays can reduce hands–on-time and the risk of sample crossover and contamination. The one-step chemistries are showing increasing use in virus detection and have been reported, in some cases, to be more appropriate than their two-step counterparts [1, 2]. Previous work presented by the Stokes Institute research group outlined a micro fluidic based continuous flow instrument which performed high throughput qPCR in nanolitre sized droplets [3]. This instrument had advantages over commercially available instruments in that it could process far more than the traditional 96 or 384 reaction setup in a single run and the reaction volume was reduced from 20–50 μl down to 30–100 nl sized droplets. Combining one-step chemistry with the technology offered by the devices being developed would lead to a high-throughput RNA-to-signal system capable of reverse transcribing and performing PCR on thousands of nanolitre sized reactions every day. It is envisaged that this technology will also lead to gene expression from single cells contained in nanolitre sized droplets. In this paper, a study was conducted in which an extra thermal region, manufactured from aluminium, was added to the existing continuous flow instruments. This region was maintained at a temperature suitable for reverse transcription, which was 48°C for the one-step kit tested. The thermal region was also a suitable length to maintain the sample at the required temperature for 15 minutes. Using a commercially available one step RT-PCR kit (TaqMan® RNA-to-CT™ 1-Step Kit, 4392653), the device was evaluated for its potential to perform one-step RT-PCR in continuously flowing nanolitre sized droplets. Electrophoresis gels were initially used in assessing specific amplification before an end-point detection method was utilized. RNA was extracted from the leukemic REH cell line with the housekeeping gene, glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) as the gene of interest. To investigate the possibility of further reducing sample preparation and facilitating further automation, amplification from cell lysates without nucleic acid extraction was carried out on the device. Cell lysates were prepared using the cell lysis buffer from the TaqMan® Gene Expression Cells-to-CT™ Kit (Cat #AM1728). It was found that the device was successful in one-step RT-PCR from extracted RNA samples and samples from cell lysates without nucleic acid extraction.
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Wirtz, Ralph M., Tilman Rau, Mark Laible, Kornelia Schlombs, Sotirios Lakis, David Wachter, Ugur Sahin, and Arndt Hartmann. "Abstract P5-10-13: Low influence of tumor cell content on mRNA expression levels of ESR, PGR, HER2 and KI67 when performing the MammaTyper® RT-PCR kit." In Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium; December 9-13, 2014; San Antonio, TX. American Association for Cancer Research, 2015. http://dx.doi.org/10.1158/1538-7445.sabcs14-p5-10-13.
Повний текст джерела
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Wallwiener, Markus, Andreas Hartkopf, Thomas Deutsch, Lakis Sotiris, Florin-Andrei Taran, Andreas Trumpp, Ralph Wirtz, and Andreas Schneeweiss. "Abstract P3-06-45: Concordance/discordance rates of HER2, ER, PR, and Ki67 in matched pair samples of primary (PBC) and metastatic breast cancer (MBC) tissues when comparing IHC with MammaTyper® RT-PCR kit." In Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium; December 9-13, 2014; San Antonio, TX. American Association for Cancer Research, 2015. http://dx.doi.org/10.1158/1538-7445.sabcs14-p3-06-45.
Повний текст джерела
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Younes, Salma, Hadeel Al-Jighefee, Farah Shurrab, Duaa Al-Sadeq, Hadi Yassine, Asmaa Althani, Reham Marei, Hashim Alhussain, and Gheyath Nasrallah. "Validation of Selected Commercial Serological Assays for Diagnosis of COVID-19." In Qatar University Annual Research Forum & Exhibition. Qatar University Press, 2020. http://dx.doi.org/10.29117/quarfe.2020.0306.
Повний текст джерелаАнотація:
As researchers around the globe rush to put the available antibody tests to use, concerns have been raised about their precision. This study aimed to evaluate and compare the performance of selected commercial & automated serological assays that are widely used in different clinical settings in Qatar. We validated the performance of five commercial IgG and IgM ELISA kits, three fully automated immunoassays, and two commercial rapid tests. The sensitivity of all assays was compared to RT-PCR and a surrogate virus neutralization test (sVNT). In addition, cross-reactivity was investigated. Among the evaluated kits, Lionex IgG assay demonstrated the best performance (~88% sensitivity and ~99 specificity). All automated assays showed an excellent correlation with the neutralization test with an overall agreement of 93.6-98.5%. The rapid assays demonstrated a very good performance in detecting IgG antibodies (86.0-88.0% sensitivity and 98.0-100% specificity).