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Phenotypic assay of a hepatitis B virus strain carrying an rtS246T variant using a new strategy. | LitMetric

Phenotypic assay of a hepatitis B virus strain carrying an rtS246T variant using a new strategy.

J Med Virol

Institute for Viral Hepatitis, Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Chongqing Medical University, Chongqing, China.

Published: January 2012

AI Article Synopsis

  • Phenotypic assays of hepatitis B virus (HBV) are crucial for studying drug resistance during long-term treatment of chronic hepatitis B.
  • A new cloning strategy using a "fragment substitution reaction" (FSR) simplifies the process of creating replication-competent HBV recombinants from clinical isolates.
  • Using this new method, researchers conducted a phenotypic assay on an HBV strain with a specific mutation, finding that it shows in vitro sensitivity to entecavir despite previously being only partially responsive.

Article Abstract

Phenotypic assays of hepatitis B virus (HBV) play an important role in research related to the problem of drug resistance that emerges during long-term nucleot(s)ide therapy in patients with chronic hepatitis B. Most of the phenotypic assay systems that are available currently rely on the transfection of recombinant replication-competent HBV DNA into hepatoma cell lines. Cloning clinical HBV isolates using conventional digestion-and-ligation techniques to generate replication-competent recombinants can be very difficult because of the sequence heterogeneity and unique structure of the HBV genome. In this study, a new strategy for constructing an HBV 1.1× recombinant was developed. The core of this strategy is the "fragment substitution reaction" (FSR). FSR allows PCR fragments to be cloned without digestion or ligation, providing a new tool for cloning fragments or genomes amplified from serum HBV DNA, and therefore making the assay of HBV phenotypes more convenient. Using this strategy, a phenotypic assay was performed on an HBV strain carrying an rtS246T variant isolated from a patient with chronic hepatitis B that was only responsive partially to entecavir therapy. The results indicated that this strain is sensitive to entecavir in vitro.

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Source
http://dx.doi.org/10.1002/jmv.22260DOI Listing

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