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. 2021 Apr 28;9(2):143-148.
doi: 10.14218/JCTH.2020.00118. Epub 2021 Mar 16.

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

Jingjing Liang  1 , Xinmiao Liang  2 , Hong Ma  1 , Leng Nie  3 , Ying Tian  4 , Guang Chen  5 , Yu Wang  1
Affiliations

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

Jingjing Liang et al. J Clin Transl Hepatol. .

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: DNA sequencing; Drug-resistance mutation; HBV DNA; Real-time PCR.

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Conflict of interest statement

The authors have no conflict of interests related to this publication.

Figures

Fig. 1
Fig. 1. Process of real-time PCR based on magnetic nanoparticles for detecting HBV DNA.
Fig. 2
Fig. 2. Amplification curves of plasmids.
Fig. 3
Fig. 3. Linear relationship between Ct values and log DNA concentrations.
See this image and copyright information in PMC

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References

    1. EASL 2017 Clinical Practice Guidelines on the management of hepatitis B virus infection. J Hepatol. 2017;67:370–398. doi: 10.1016/j.jhep.2017.03.021. - DOI - PubMed
    1. Hermans LE, Svicher V, Pas SD, Salpini R, Alvarez M, Ben Ari Z, et al. Combined analysis of the prevalence of drug-resistant hepatitis B virus in antiviral therapy-experienced patients in Europe (CAPRE) J Infect Dis. 2016;213:39–48. doi: 10.1093/infdis/jiv363. - DOI - PubMed
    1. Terrault NA, Bzowej NH, Chang KM, Hwang JP, Jonas MM, Murad MH. AASLD guidelines for treatment of chronic hepatitis B. Hepatology. 2016;63:261–283. doi: 10.1002/hep.28156. - DOI - PMC - PubMed
    1. Lin CL, Kao JH. Natural history of acute and chronic hepatitis B: The role of HBV genotypes and mutants. Best Pract Res Clin Gastroenterol. 2017;31:249–255. doi: 10.1016/j.bpg.2017.04.010. - DOI - PubMed
    1. Wang W, Shu Y, Bao H, Zhao W, Wang W, Wang Q, et al. Genotypes and hot spot mutations of hepatitis B virus in northwest Chinese population and its correlation with diseases progression. Biomed Res Int. 2019;2019:3890962. doi: 10.1155/2019/3890962. - DOI - PMC - PubMed

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