A novel method for the analysis of drug-resistant phenotypes of hepatitis B virus

  • Authors:
    • Bo Qin
    • Tingting He
    • Zhijun Chen
    • Wenying Xu
    • Guoshao Pan
    • Chunyu Tu
  • View Affiliations

  • Published online on: February 11, 2013     https://doi.org/10.3892/ijmm.2013.1277
  • Pages: 975-981
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Abstract

Chronic hepatitis B virus (CHB) infection is a major cause of cirrhosis and hepatocellular carcinoma. Nucleoside analogs (NAs) are popularly used to treat chronic hepatitis B virus (HBV) infections; however, the anti-HBV effect is attenuated by drug-resistant viral mutations selected during long-term antiviral therapy. The timely analysis of drug-resistance mutations is essential in order to adjust treatment regimes. In this study, a T1699C substitution was introduced into the x gene of pHBV1.3 to generate an additional XhoI site, termed pHBV1.3‑XhoI, which is a nonsense mutation and does not influence protein expression, HBV replication ability, or NA susceptibility. Based on co-transfection with weak or non-replicative HBV plasmids and pHBV1.3-XhoI or pHBV1.3 and -XhoI-P-null plasmids into hepatocellular carcinoma cells, PCR was used to amplify 1176‑bp segments of T/C1699 using the isolated HBV encapsulated DNA as a template, modified by XhoI digestion and subjected to agarose gel electrophoresis. Different bands composed of different virions were used to distinguish the replication capacities of the plasmids. Our results demonstrated no significant effects when different virions co-existed. A novel resistance test method was developed by co-transfection with pHBV1.3-XhoI and -rtL180M/M204V and treatment with various NA concentrations. Different bands composed of pHBV1.3-XhoI or -rtL180M/M204V were used to distinguish NA susceptibility. The bands composed of pHBV1.3 were more sharply reduced by lamivudine (LMV) than -rtL180M/M204V. The data demonstrate that the method established in our study may be used for the analysis of drug-resistant phenotypes at the cellular level.

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April 2013
Volume 31 Issue 4

Print ISSN: 1107-3756
Online ISSN:1791-244X

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Spandidos Publications style
Qin B, He T, Chen Z, Xu W, Pan G and Tu C: A novel method for the analysis of drug-resistant phenotypes of hepatitis B virus. Int J Mol Med 31: 975-981, 2013.
APA
Qin, B., He, T., Chen, Z., Xu, W., Pan, G., & Tu, C. (2013). A novel method for the analysis of drug-resistant phenotypes of hepatitis B virus. International Journal of Molecular Medicine, 31, 975-981. https://doi.org/10.3892/ijmm.2013.1277
MLA
Qin, B., He, T., Chen, Z., Xu, W., Pan, G., Tu, C."A novel method for the analysis of drug-resistant phenotypes of hepatitis B virus". International Journal of Molecular Medicine 31.4 (2013): 975-981.
Chicago
Qin, B., He, T., Chen, Z., Xu, W., Pan, G., Tu, C."A novel method for the analysis of drug-resistant phenotypes of hepatitis B virus". International Journal of Molecular Medicine 31, no. 4 (2013): 975-981. https://doi.org/10.3892/ijmm.2013.1277