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Betaine‐assisted recombinase polymerase assay for rapid hepatitis B virus detection
Hepatitis B virus (HBV) is a worldwide epidemic pathogen that causes hepatitis B. On‐site screening the HBV infection is of critical importance for preventing and diagnosing HBV infection. In this paper, a simple, visual, and rapid method for on‐site detection of HBV‐DNA has been developed. This met...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8411421/ https://www.ncbi.nlm.nih.gov/pubmed/32388885 http://dx.doi.org/10.1002/bab.1940 |
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author | Yi, Ting‐ting Zhang, Han‐yun Liang, Hua Gong, Guo‐zhong Cai, Yan |
author_facet | Yi, Ting‐ting Zhang, Han‐yun Liang, Hua Gong, Guo‐zhong Cai, Yan |
author_sort | Yi, Ting‐ting |
collection | PubMed |
description | Hepatitis B virus (HBV) is a worldwide epidemic pathogen that causes hepatitis B. On‐site screening the HBV infection is of critical importance for preventing and diagnosing HBV infection. In this paper, a simple, visual, and rapid method for on‐site detection of HBV‐DNA has been developed. This method is based on betaine‐assisted recombinase polymerase assay and followed with naked‐eye detection via lateral flow assay (BRPA‐LF). Result show that nonspecific amplification is prone to occur in recombinase polymerase amplification (RPA) if the assay was performed with serum sample without purification. This problem has been addressed by adding 0.8 M of betaine to the RPA reactions. It was demonstrated that BRPA‐LF can detect 1,000 copies of HBV‐DNA in 50 μL mixture, and achieved 90% sensitivity and 100% specificity for serum sample detection. These results demonstrated that BRPA‐LF can resist serum interference and has great potential for on‐site screening of HBV infection. |
format | Online Article Text |
id | pubmed-8411421 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-84114212021-09-08 Betaine‐assisted recombinase polymerase assay for rapid hepatitis B virus detection Yi, Ting‐ting Zhang, Han‐yun Liang, Hua Gong, Guo‐zhong Cai, Yan Biotechnol Appl Biochem Original Articles Hepatitis B virus (HBV) is a worldwide epidemic pathogen that causes hepatitis B. On‐site screening the HBV infection is of critical importance for preventing and diagnosing HBV infection. In this paper, a simple, visual, and rapid method for on‐site detection of HBV‐DNA has been developed. This method is based on betaine‐assisted recombinase polymerase assay and followed with naked‐eye detection via lateral flow assay (BRPA‐LF). Result show that nonspecific amplification is prone to occur in recombinase polymerase amplification (RPA) if the assay was performed with serum sample without purification. This problem has been addressed by adding 0.8 M of betaine to the RPA reactions. It was demonstrated that BRPA‐LF can detect 1,000 copies of HBV‐DNA in 50 μL mixture, and achieved 90% sensitivity and 100% specificity for serum sample detection. These results demonstrated that BRPA‐LF can resist serum interference and has great potential for on‐site screening of HBV infection. John Wiley and Sons Inc. 2020-06-30 2021-06 /pmc/articles/PMC8411421/ /pubmed/32388885 http://dx.doi.org/10.1002/bab.1940 Text en © 2020 Affiliated Hospital of North Sichuan Medical College. Biotechnology and Applied Biochemistry published by Wiley Periodicals LLC on behalf of International Union of Biochemistry and Molecular Biology https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Articles Yi, Ting‐ting Zhang, Han‐yun Liang, Hua Gong, Guo‐zhong Cai, Yan Betaine‐assisted recombinase polymerase assay for rapid hepatitis B virus detection |
title | Betaine‐assisted recombinase polymerase assay for rapid hepatitis B virus detection |
title_full | Betaine‐assisted recombinase polymerase assay for rapid hepatitis B virus detection |
title_fullStr | Betaine‐assisted recombinase polymerase assay for rapid hepatitis B virus detection |
title_full_unstemmed | Betaine‐assisted recombinase polymerase assay for rapid hepatitis B virus detection |
title_short | Betaine‐assisted recombinase polymerase assay for rapid hepatitis B virus detection |
title_sort | betaine‐assisted recombinase polymerase assay for rapid hepatitis b virus detection |
topic | Original Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8411421/ https://www.ncbi.nlm.nih.gov/pubmed/32388885 http://dx.doi.org/10.1002/bab.1940 |
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