乙型肝炎病毒基因聚合酶链反应分型方法的建立

根据乙型肝炎病毒（ＨＢＶ）的Ｓ区序列设计了３条引物：ＨＢＳ１、ＨＢＳ２和ＨＢＳ３，与ＨＢＳ１和ＨＢＳ２配对，经２次聚合酶链反应（ＰＣＲ）扩增。即可在检测有无ＨＢＶ－ＤＮＡ存在的同时对其基因型分类，可检出１０ａｇ的ＨＢＶ－ＤＮＡ，比免疫学方法更灵敏和特异。２５份不同亚型的标准血清中的绝大多数用Ｓ－ＰＣＲ和ＡＧＩＤ／ＲＰＨＡ的分型结果一致，Ｓ－ＰＣＲ的另一突出优越性的于能够对ＨＢｓＡｇ低滴度和阴性标本

Typing of Hepatitis B Virus Gene by Polymerase Chain Reaction

Three oligonucleotide primers were selected by the S-PCR method according to S gene sequence of adr subtype;HBS1;allowing HBV DNA amplification of adr and ayr subtypes but not that of adw and ayw subtypes,and HBS2;adr and adw but not ayr and ayw.The HBS3 is a reverse (antisense) primer that was paired with the HBS1 and HBS2 primers.Thus,the HBV genotype could be easily identified by two steps of PCR amplification.The sensitivity of S-PCR was 10ag of HBV DNA,much higher than that of immunological subtyping methods as well as more specific and valid.The results of S-PCR were identical to those obtained by AGID and RPHA with monoclonal antibodies in 25 standard serum samples of different subtypes.Another prominant advantage of S-PCR is that it allows subtyping of HBV in the serum of those without HBsAg or containing HBsAg in concentrations too low to be subtyped by conventional immunological methods.