Phenotypic assay of a hepatitis B virus strain carrying an rtS246T variant using a new strategy.
J Med Virol
; 84(1): 34-43, 2012 Jan.
Article
en En
| MEDLINE
| ID: mdl-22052677
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.
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Antivirales
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Virología
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Virus de la Hepatitis B
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ADN Polimerasa Dirigida por ARN
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Clonación Molecular
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Mutación Missense
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Farmacorresistencia Viral
Límite:
Adult
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Humans
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Male
Idioma:
En
Revista:
J Med Virol
Año:
2012
Tipo del documento:
Article
País de afiliación:
China