YMDD variants of HBV DNA polymerase gene: rapid detection and clinicopathological analysis with long-term lamivudine therapy after liver transplantation

乙型肝炎病毒DNA聚合酶基因YMDD变异株:肝移植后长期拉米夫定治疗的快速检测及临床病理分析

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作者:Fei Pei,Jun-Yu Ning, Jiang-Feng You, Jing-Pin Yang, Jie Zheng

Abstract

Aim: To look for a rapid low-cost technique for the detection of HBV variants. Methods: Two patients who underwent orthotopic liver transplantation (OLT) for HBV infection were treated with lamivudine (100 mg daily) and HBV infection recurred in the grafted livers. The patients were monitored intensively for liver enzymes, hepatitis B surface antigen (HBsAg) and HBV DNA in serum. Liver biopsy was performed regularly. HBV DNA in a conserved polymerase domain (the YMDD locus) was amplified from serum of each patient by PCR and sequenced. HBV genotypes were analyzed by restriction fragment length polymorphism (RFLP) of the PCR products generated from a fragment of the polymerase gene. Results: YMDD wild-type HBV was detected in one patient by PCR-RFLP and DNA sequencing 19 mo after OLT, and YIDD mutant-type HBV in the other patient, 16 mo after OLT. Conclusion: PCR-RFLP assay is an accurate and simple method for genotyping lamivudine-resistant HBV variants.

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