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Simultaneous detection of Marburg virus and Ebola virus with TaqMan‐based multiplex real‐time PCR method

BACKGROUND: Marburg virus (MARV) and Ebola virus (EBOV) are acute infections with high case fatality rates. It is of great significance for epidemic monitoring and prevention and control of infectious diseases by the development of a rapid, specific, and sensitive quantitative PCR method to detect t...

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Autores principales: Yu, Zhikang, Wu, Heming, Huang, Qingyan, Zhong, Zhixiong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8183904/
https://www.ncbi.nlm.nih.gov/pubmed/33939238
http://dx.doi.org/10.1002/jcla.23786
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author Yu, Zhikang
Wu, Heming
Huang, Qingyan
Zhong, Zhixiong
author_facet Yu, Zhikang
Wu, Heming
Huang, Qingyan
Zhong, Zhixiong
author_sort Yu, Zhikang
collection PubMed
description BACKGROUND: Marburg virus (MARV) and Ebola virus (EBOV) are acute infections with high case fatality rates. It is of great significance for epidemic monitoring and prevention and control of infectious diseases by the development of a rapid, specific, and sensitive quantitative PCR method to detect two pathogens simultaneously. METHODS: Primers and TaqMan probes were designed according to highly conserved sequences of these viruses. Sensitivity, specificity, linear range, limit of detection, and the effects of hemolysis and lipid on real‐time qPCR were evaluated. RESULTS: The linearity of the curve allowed quantification of nucleic acid concentrations in range from 10(3) to 10(9) copies/ml per reaction (MARV and EBOV). The limit of detection of EBOV was 40 copies/ml, and MARV was 100 copies/ml. It has no cross‐reaction with other pathogens such as hepatitis b virus (HBV), hepatitis c virus (HCV), human papillomavirus (HPV), Epstein‐Barr virus (EBV), herpes simplex virus (HSV), cytomegalovirus (CMV), and human immunodeficiency virus (HIV). Repeatability analysis of the two viruses showed that their coefficient of variation (CV) was less than 5.0%. The above results indicated that fluorescence quantitative PCR could detect EBOV and MARV sensitively and specifically. CONCLUSIONS: The TaqMan probe‐based multiplex fluorescence quantitative PCR assays could detect EBOV and MARV sensitively specifically and simultaneously.
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spelling pubmed-81839042021-06-16 Simultaneous detection of Marburg virus and Ebola virus with TaqMan‐based multiplex real‐time PCR method Yu, Zhikang Wu, Heming Huang, Qingyan Zhong, Zhixiong J Clin Lab Anal Research Articles BACKGROUND: Marburg virus (MARV) and Ebola virus (EBOV) are acute infections with high case fatality rates. It is of great significance for epidemic monitoring and prevention and control of infectious diseases by the development of a rapid, specific, and sensitive quantitative PCR method to detect two pathogens simultaneously. METHODS: Primers and TaqMan probes were designed according to highly conserved sequences of these viruses. Sensitivity, specificity, linear range, limit of detection, and the effects of hemolysis and lipid on real‐time qPCR were evaluated. RESULTS: The linearity of the curve allowed quantification of nucleic acid concentrations in range from 10(3) to 10(9) copies/ml per reaction (MARV and EBOV). The limit of detection of EBOV was 40 copies/ml, and MARV was 100 copies/ml. It has no cross‐reaction with other pathogens such as hepatitis b virus (HBV), hepatitis c virus (HCV), human papillomavirus (HPV), Epstein‐Barr virus (EBV), herpes simplex virus (HSV), cytomegalovirus (CMV), and human immunodeficiency virus (HIV). Repeatability analysis of the two viruses showed that their coefficient of variation (CV) was less than 5.0%. The above results indicated that fluorescence quantitative PCR could detect EBOV and MARV sensitively and specifically. CONCLUSIONS: The TaqMan probe‐based multiplex fluorescence quantitative PCR assays could detect EBOV and MARV sensitively specifically and simultaneously. John Wiley and Sons Inc. 2021-05-03 /pmc/articles/PMC8183904/ /pubmed/33939238 http://dx.doi.org/10.1002/jcla.23786 Text en © 2021 The Authors. Journal of Clinical Laboratory Analysis published by Wiley Periodicals LLC https://creativecommons.org/licenses/by-nc/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ (https://creativecommons.org/licenses/by-nc/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.
spellingShingle Research Articles
Yu, Zhikang
Wu, Heming
Huang, Qingyan
Zhong, Zhixiong
Simultaneous detection of Marburg virus and Ebola virus with TaqMan‐based multiplex real‐time PCR method
title Simultaneous detection of Marburg virus and Ebola virus with TaqMan‐based multiplex real‐time PCR method
title_full Simultaneous detection of Marburg virus and Ebola virus with TaqMan‐based multiplex real‐time PCR method
title_fullStr Simultaneous detection of Marburg virus and Ebola virus with TaqMan‐based multiplex real‐time PCR method
title_full_unstemmed Simultaneous detection of Marburg virus and Ebola virus with TaqMan‐based multiplex real‐time PCR method
title_short Simultaneous detection of Marburg virus and Ebola virus with TaqMan‐based multiplex real‐time PCR method
title_sort simultaneous detection of marburg virus and ebola virus with taqman‐based multiplex real‐time pcr method
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8183904/
https://www.ncbi.nlm.nih.gov/pubmed/33939238
http://dx.doi.org/10.1002/jcla.23786
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