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Detection of Hepatitis B Virus M204V Mutation Quantitatively via Real-time PCR
BACKGROUND AND AIMS: Drug-resistant DNA mutations of the hepatitis B virus (HBV) affect treatment response in chronic hepatitis B patients. We have established a new, sensitive, specific, accurate and convenient real-time PCR method to detect HBV mutations quantitatively. METHODS: Blood samples were...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
XIA & HE Publishing Inc.
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8111100/ https://www.ncbi.nlm.nih.gov/pubmed/34007795 http://dx.doi.org/10.14218/JCTH.2020.00118 |
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author | Liang, Jingjing Liang, Xinmiao Ma, Hong Nie, Leng Tian, Ying Chen, Guang Wang, Yu |
author_facet | Liang, Jingjing Liang, Xinmiao Ma, Hong Nie, Leng Tian, Ying Chen, Guang Wang, Yu |
author_sort | Liang, Jingjing |
collection | PubMed |
description | BACKGROUND AND AIMS: Drug-resistant DNA mutations of the hepatitis B virus (HBV) affect treatment response in chronic hepatitis B patients. We have established a new, sensitive, specific, accurate and convenient real-time PCR method to detect HBV mutations quantitatively. METHODS: Blood samples were collected from patients showing viral breakthrough, primary nonresponse, or poor response during treatment, and mutations were detected via direct sequencing to assess our method. A plasmid containing the M204V mutation was synthesized and standard curves plotted. RESULTS: The determination coefficient for linear correlation between Ct and log plasmid copy numbers was 0.996, where Ct value was −3.723log (DNA concentration) +48.647. Coefficients of variation indicated good reproducibility. Correctness was within tolerable bias. Limit of detection was 10(3) copies/mL. Specificity, accuracy, positive predictive value and negative predictive value were 92.86%, 100%, 96.88%, 100% and 94.74%, respectively. CONCLUSIONS: These results show that our method can be used to detect HBV M204V mutations with the advantages of sensitivity, specificity and efficiency, providing a new choice for monitoring drug resistance. |
format | Online Article Text |
id | pubmed-8111100 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | XIA & HE Publishing Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-81111002021-05-17 Detection of Hepatitis B Virus M204V Mutation Quantitatively via Real-time PCR Liang, Jingjing Liang, Xinmiao Ma, Hong Nie, Leng Tian, Ying Chen, Guang Wang, Yu J Clin Transl Hepatol Original Article BACKGROUND AND AIMS: Drug-resistant DNA mutations of the hepatitis B virus (HBV) affect treatment response in chronic hepatitis B patients. We have established a new, sensitive, specific, accurate and convenient real-time PCR method to detect HBV mutations quantitatively. METHODS: Blood samples were collected from patients showing viral breakthrough, primary nonresponse, or poor response during treatment, and mutations were detected via direct sequencing to assess our method. A plasmid containing the M204V mutation was synthesized and standard curves plotted. RESULTS: The determination coefficient for linear correlation between Ct and log plasmid copy numbers was 0.996, where Ct value was −3.723log (DNA concentration) +48.647. Coefficients of variation indicated good reproducibility. Correctness was within tolerable bias. Limit of detection was 10(3) copies/mL. Specificity, accuracy, positive predictive value and negative predictive value were 92.86%, 100%, 96.88%, 100% and 94.74%, respectively. CONCLUSIONS: These results show that our method can be used to detect HBV M204V mutations with the advantages of sensitivity, specificity and efficiency, providing a new choice for monitoring drug resistance. XIA & HE Publishing Inc. 2021-04-28 2021-03-16 /pmc/articles/PMC8111100/ /pubmed/34007795 http://dx.doi.org/10.14218/JCTH.2020.00118 Text en © 2021 Authors. https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution-Noncommercial 4.0 International License (CC BY-NC 4.0), permitting all non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Article Liang, Jingjing Liang, Xinmiao Ma, Hong Nie, Leng Tian, Ying Chen, Guang Wang, Yu Detection of Hepatitis B Virus M204V Mutation Quantitatively via Real-time PCR |
title | Detection of Hepatitis B Virus M204V Mutation Quantitatively via Real-time PCR |
title_full | Detection of Hepatitis B Virus M204V Mutation Quantitatively via Real-time PCR |
title_fullStr | Detection of Hepatitis B Virus M204V Mutation Quantitatively via Real-time PCR |
title_full_unstemmed | Detection of Hepatitis B Virus M204V Mutation Quantitatively via Real-time PCR |
title_short | Detection of Hepatitis B Virus M204V Mutation Quantitatively via Real-time PCR |
title_sort | detection of hepatitis b virus m204v mutation quantitatively via real-time pcr |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8111100/ https://www.ncbi.nlm.nih.gov/pubmed/34007795 http://dx.doi.org/10.14218/JCTH.2020.00118 |
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