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作 者:杨洁[1] 骆抗先[1] 郭亚兵[1] 戴琳[1] 闫丽[1] 候金林[1]
机构地区:[1]第一军医大学南方医院感染内科,广州510515
出 处:《中华肝脏病杂志》2002年第1期55-57,共3页Chinese Journal of Hepatology
基 金:国家自然科学基金(3980012);广东省自然科学基金(980223)
摘 要:目的 建立一种简便易行的乙型肝炎病毒基因型分型方法(只需 PCR)。方法 对 GenBank中查获的114例HBV全序列进行比较分析,找出每种基因型相对于其他5种基因型的独特序列,并根据这些独特序列设计出6对分别针对A-F基因型的特异引物。用这6对引物分别对标本进行PCR,根据阳性结果判断出标本的基因型。进一步简化该方法,用多引物对PCR法(PCR with several primer sets in a single tube,Multiplex PCR)将B、C、D 3种基因型的特异引物混台进行PCR,根据扩增片断的大小判断基因型。用此方祛对已鉴定的B、C、D型标本进行比较和验证。结果 单引物对PCR与多引物对PCR的分型结果一致,且与以前用PCR-RFLP法的分型结果一致。结论 用多引物对PCR分型法准确易行,灵敏性高,便于推广应用。Objective To establish a simple and practicable method to identify the different genotypes of hepatitis B virus (HBV). Methods Based on the alignment of 114 complete nucleotide sequence for HBV DNA of different genotypes, the specific sequence of each genotype was found. Six primer sets were designed for each of the six genotypes according to the genotype-specific sequence, and used separately for PCR. The genotype of HBV was identified according to the positive result of PCR. Three primer sets for B, C and D genotypes were added into a single tube for PCR reaction, and HBV was genotyped according to the length of the amplified DNA. Results There was no difference in the genotyping result of PCR by single or multiplex primers. which was identical to the PCR-RFLP method. Conclusions This multiplex PCR method is simple, precise. sensitive, and easy to use.
关 键 词:乙型肝炎病毒 基因型 聚合酶链反应 PCR 多引物对 分型 A-F
分 类 号:R373.21[医药卫生—病原生物学]
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