Journal of Clinical and Translational Hepatology

Journal of Clinical and Translational Hepatology

Friday, 05 / 14 / 2021

Articles

Abstract

ORIGINAL ARTICLE

Detection of Hepatitis B Virus M204V Mutation Quantitatively via Real-time PCR

Jingjing Liang1, Xinmiao Liang3, Hong Ma1, Leng Nie4, Ying Tian5Guang Chenand Yu Wang1,*

1  Liver Research Center, Beijing Friendship Hospital, Capital Medical University, National Clinical Research Center of Digestive Diseases, Beijing, China
2  Technical center of Beijing Customs District, Beijing, China
3  Suzhou Primreg Gene Company, Suzhou, China
4  Department of Geriatrics, Beijing Friendship Hospital, Capital Medical University, Beijing, China
5  Department of Interventional Radiology, Beijing Friendship Hospital, Capital Medical University, Beijing, China
*Correspondence to: Guang Chen, Department of Interventional Radiology, Beijing Friendship Hospital, Capital Medical University, 95 Yong-an Road, Xicheng District, Beijing 100050, China. Tel: +86-131-4126-5625, E-mail: This email address is being protected from spambots. You need JavaScript enabled to view it. ;Yu Wang, Liver Research Center, Beijing Friendship Hospital, Capital Medical University, National Clinical Research Center of Digestive Diseases, 95 Yong-an Road, Xicheng District, Beijing 100050, China. Tel: +86-133-1110-0797, E-mail:  This email address is being protected from spambots. You need JavaScript enabled to view it.

Journal of Clinical and Translational Hepatology 2021;9(2):143-148 DOI: 10.14218/JCTH.2020.00118
Received: November 15, 2020 Accepted: January 30, 2021 Published online: March 16, 2021

Abstract

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 103 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.

Keywords

HBV DNA, Drug-resistance mutation, Real-time PCR, DNA sequencing

Journal of Clinical and Translational Hepatology 2021 vol. 9, 143-148  [ Html  ] [ PDF Full-text ]

© 2021 Authors. This is an Open Access article distributed under the terms of the  Creative Commons Attribution-Noncommercial 4.0 License(CC BY-NC 4.0), permitting all non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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