摘要
目的 建立乙型肝炎病毒 (HBV)S基因聚合酶链反应 限制性片段长度多态性 (PCR RFLP)的基因型分型方法并验证其准确性。方法 比较GenBank中HBV 2 2 3株各基因型全序列的S基因氨基酸及核苷酸序列 ,设计限制性内切酶BsrI、StyI、DpnI和HpaII鉴别HBVA F基因型的方法。选择经前SPCR PFLP的方法已鉴定出的HBVB、C和D基因型标本各 3 0例 ,用此方法鉴定验证 ,并随机对HBVB、C和D基因型各 3株标本进行S基因测序证实。结果 两种PCR RFLP及S基因测序鉴定HBV基因型结果一致。结论 S基因PCR RFLP的方法能够准确鉴定HBV基因型。此方法灵敏性高 ,特异性强 ,且酶切图谱简明单一 ,易于识别 ,适宜于HBV基因型的流行病学调查。
Objective A method was established for genotyping of hepatitis B virus (HBV),based on the restriction fragment length polymorphism(RFLP)created by BsrI, StyI,DpnI and HpaII action on an amplified segment of the S region. Methods 223 full-genomic sequences were analyzed and the aligned nucleotide and amino acid sequences of S gene, genotype specific regions were identified by the restriction enzymes, BsrI, StyI,DpnI and HpaII. Pre S PCR-RFLP genotyping method was applied to a number of serum samples from hepatitis B e antigen (HBeAg) positive and negative Chinese chronic HBV carriers. And in 90 samples the following genotypes were observed: 30B, 30 C, 30D. The method using S gene PCR-RFLP was confirmed to be correct by these 90 samples. Three samples of each genotype B, C and D were randomly selected and directly sequenced their S gene to confirm that HBV S gene PCR-RFLP genotyping method was correct disectly. Results The results of two PCR-RFLP HBV genotyping methods were coincide with that of S gene sequence. Conclusions The method for genotyping of hepatitis B virus (HBV), based on S gene RFLP is established to be highly sensitive, differential and accurate. The RFLP patterns are easy to be recognized because of its simplicity and singleness.
出处
《中华传染病杂志》
CAS
CSCD
北大核心
2001年第4期224-228,共5页
Chinese Journal of Infectious Diseases
基金
国家自然科学基金资助项目 ( 39630 2 80 )
