| Summary: | 碩士 === 國立成功大學 === 分子醫學研究所 === 92 === Chronic infection of hepatitis B virus (HBV) is considered as an increasing risk factor developing cirrhosis and hepatocellular carcinoma. In the treatment of chronic hepatitis B patients, the use of lamivudine is effective to suppress viral replication; however, durable suppression of viral replication is limited by development of drug-resistant strains upon prolonged therapy. In the present study, we aimed to analyze the influences of virological factors involved in the viral fitness during antiviral therapies, and develop sensitive methods for early detections of HBV YMDD mutations. Several primers and restriction enzyme cutting sites were designed for the detection of the appearance of viral mutants. The PCR-enriched RFLP results showed that strategies for eliminating the co-existed wild type viruses were effective, and the detecting sensitivity were enhanced by about 100 folds. The sensitivity of this method is evaluated to 101 copies/ml, with identical results between nucleotide sequence analyses. This method is thus effective for early detection of the developing lamivudine-resistant mutants, and monitoring of the entire antiviral therapies. Despite for the fact that only incomplete understanding of the factors involved in the viral-host interactions, this method is valuable and useful for the detecting of drug-resistant mutants for clinical implications.
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