Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Yi, Ting‐ting, Zhang, Han‐yun, Liang, Hua, Gong, Guo‐zhong, Cai, Yan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2020
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
_version_ 1783747293526097920
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
work_keys_str_mv AT yitingting betaineassistedrecombinasepolymeraseassayforrapidhepatitisbvirusdetection
AT zhanghanyun betaineassistedrecombinasepolymeraseassayforrapidhepatitisbvirusdetection
AT lianghua betaineassistedrecombinasepolymeraseassayforrapidhepatitisbvirusdetection
AT gongguozhong betaineassistedrecombinasepolymeraseassayforrapidhepatitisbvirusdetection
AT caiyan betaineassistedrecombinasepolymeraseassayforrapidhepatitisbvirusdetection