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目的探讨基因分型结合克隆后测序方法在动态监测ＨＢＶ基因变异中的价值。方法分析１例重型肝炎患者发 病后血清，用 ＰＣＲ扩增血清中 ＨＢＶ ｐｒｅＳ／Ｓ基因片段并克隆，２份标本各随机挑取 ３个阳性克隆进行测序；同时用 ＲＦＬＰ方法分析２份血清中ＨＢＶ毒株的基因型。结果ＰＦＬＰ分析２份血清为Ｂ基因型为主的Ｂ／Ｃ基因型混合毒株感 染；６个克隆中仅１株为Ｃ基因型。比较２份血清ＨＢＶ ｐｒｅＳ／Ｓ克隆核酸序列，有１７０个点位的差异，剔除Ｃ基因型株 后，仅５４个位点差异。结论克隆后测序结合ＲＦＬＰ对ＨＢＶ基因型测定有助于更准确地对ＨＢＶ基因变异进行动态分 析。
Objective To investigate the value of genotyping combined with post-cloning sequencing in the dynamic monitoring of HBV gene mutation. Methods Serum samples were collected from one patient with severe hepatitis. PCR was used to amplify the HBV preS / S gene fragment in the serum and cloned. Two positive clones were randomly selected from each of the two samples and sequenced. In the meanwhile, Genotype of strain. Results PFLP analysis showed that two of the sera were infected with the B / C genotype mixed with B genotype. Only one of the six clones was genotype C. Comparing the two serum HBV preS / S clone nucleic acid sequences, there were 170 loci differences, excluding only 54 loci after C genotype was removed. Conclusion The clonal sequencing combined with RFLP can be used to analyze HBV genotypes more accurately.