摘要
根据乙型肝炎病毒（ＨＢＶ）的Ｓ区序列设计了３条引物：ＨＢＳ１（ｗ－／ｒ＋）、ＨＢＳ２（ｄ＋／ｙ－）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的另一突出优越性在于能够对ＨＢｓＡｇ低滴度和阴性标本分型。
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.
出处
《中华实验和临床病毒学杂志》
CAS
CSCD
1995年第1期29-32,共4页
Chinese Journal of Experimental and Clinical Virology
关键词
基因
聚合酶链反应
方法
乙型肝炎病毒
Subtypes of hepatitis B virus
Hepatitis B virus DNA
Polymerase chain reaction
Subtypes of HBsAg
