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Rapid two‐stage amplification in a single tube for simultaneous detection of norovirus GII and group a rotavirus
The most prevalent viruses currently causing diarrhea are norovirus and rotavirus, and rapid and sensitive detection methods are essential for the early diagnosis of disease. The purpose of this study was to establish a sensitive single‐tube two‐stage nucleic acid amplification method—reverse transc...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10098066/ https://www.ncbi.nlm.nih.gov/pubmed/36916770 http://dx.doi.org/10.1002/jcla.24858 |
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author | Li, Feng‐yu Guo, Ying‐hui Sun, Zhen‐lu Liu, Hong Zhao, Meng‐chuan Cui, Jia Jiang, Yue Shen, Xin‐xin Ma, Xue‐jun Feng, Zhi‐shan |
author_facet | Li, Feng‐yu Guo, Ying‐hui Sun, Zhen‐lu Liu, Hong Zhao, Meng‐chuan Cui, Jia Jiang, Yue Shen, Xin‐xin Ma, Xue‐jun Feng, Zhi‐shan |
author_sort | Li, Feng‐yu |
collection | PubMed |
description | The most prevalent viruses currently causing diarrhea are norovirus and rotavirus, and rapid and sensitive detection methods are essential for the early diagnosis of disease. The purpose of this study was to establish a sensitive single‐tube two‐stage nucleic acid amplification method—reverse transcription recombinase‐assisted PCR (RT‐RAP)—for simultaneous detection of norovirus GII and group A Rotavirus, with the first stage consisting of isothermal reverse transcription recombinase‐aided amplification (RT‐RAA) and the second stage consisting of qPCR (quantitative PCR). RT‐RAP is more sensitive than either RT‐RAA or qRT‐PCR (quantitative RT‐PCR) alone. And the addition of a barrier that can be disassembled after heating enabled the detection of samples within 1 h in a single closed tube. Sensitivity was 10 copies/reaction of norovirus (Novs) GII and group A rotavirus (RVA). In parallel, two hundred fecal specimens were used to evaluate the method and compare it with a commercial fluorescent quantitative RT‐PCR. The data showed kappa values of 0.957 and 0.98 (p < 0.05) for detecting Novs GII and RVA by the two methods, indicating the potential of the newly established assay to be applied to clinical and laboratory testing. |
format | Online Article Text |
id | pubmed-10098066 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-100980662023-04-14 Rapid two‐stage amplification in a single tube for simultaneous detection of norovirus GII and group a rotavirus Li, Feng‐yu Guo, Ying‐hui Sun, Zhen‐lu Liu, Hong Zhao, Meng‐chuan Cui, Jia Jiang, Yue Shen, Xin‐xin Ma, Xue‐jun Feng, Zhi‐shan J Clin Lab Anal Research Articles The most prevalent viruses currently causing diarrhea are norovirus and rotavirus, and rapid and sensitive detection methods are essential for the early diagnosis of disease. The purpose of this study was to establish a sensitive single‐tube two‐stage nucleic acid amplification method—reverse transcription recombinase‐assisted PCR (RT‐RAP)—for simultaneous detection of norovirus GII and group A Rotavirus, with the first stage consisting of isothermal reverse transcription recombinase‐aided amplification (RT‐RAA) and the second stage consisting of qPCR (quantitative PCR). RT‐RAP is more sensitive than either RT‐RAA or qRT‐PCR (quantitative RT‐PCR) alone. And the addition of a barrier that can be disassembled after heating enabled the detection of samples within 1 h in a single closed tube. Sensitivity was 10 copies/reaction of norovirus (Novs) GII and group A rotavirus (RVA). In parallel, two hundred fecal specimens were used to evaluate the method and compare it with a commercial fluorescent quantitative RT‐PCR. The data showed kappa values of 0.957 and 0.98 (p < 0.05) for detecting Novs GII and RVA by the two methods, indicating the potential of the newly established assay to be applied to clinical and laboratory testing. John Wiley and Sons Inc. 2023-03-14 /pmc/articles/PMC10098066/ /pubmed/36916770 http://dx.doi.org/10.1002/jcla.24858 Text en © 2023 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 Li, Feng‐yu Guo, Ying‐hui Sun, Zhen‐lu Liu, Hong Zhao, Meng‐chuan Cui, Jia Jiang, Yue Shen, Xin‐xin Ma, Xue‐jun Feng, Zhi‐shan Rapid two‐stage amplification in a single tube for simultaneous detection of norovirus GII and group a rotavirus |
title | Rapid two‐stage amplification in a single tube for simultaneous detection of norovirus GII and group a rotavirus |
title_full | Rapid two‐stage amplification in a single tube for simultaneous detection of norovirus GII and group a rotavirus |
title_fullStr | Rapid two‐stage amplification in a single tube for simultaneous detection of norovirus GII and group a rotavirus |
title_full_unstemmed | Rapid two‐stage amplification in a single tube for simultaneous detection of norovirus GII and group a rotavirus |
title_short | Rapid two‐stage amplification in a single tube for simultaneous detection of norovirus GII and group a rotavirus |
title_sort | rapid two‐stage amplification in a single tube for simultaneous detection of norovirus gii and group a rotavirus |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10098066/ https://www.ncbi.nlm.nih.gov/pubmed/36916770 http://dx.doi.org/10.1002/jcla.24858 |
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