[目的]建立一种在临床上准确、快速、简便地对乙型肝炎病毒进行基因分型(BCD三型)的方法.[方法]大量分析已分型的HBV基因组序列,寻找出HBV各基因型的型特异性碱基的分布规律,设计型特异性的引物和探针,利用已知全序列HBV建立荧光PCR的基因分型方法.并与PCR-RFLP及S基因测序等分型方法作比较.[结果]荧光PCR方法对已知基因组序列的HBV分型结果与基因组分析完全一致,利用大量随机临床标本摸索的最佳反应条件为94℃,1 min;94℃,10 s,60℃,30 s,40个循环.与PCR-RFLP相比,分型结果一致性在84.0%;与S基因测序分型相比,一致性为96.2%.[结论]利用荧光PCR能灵敏、快速、准确地进行乙型肝炎病毒基因分型,方便临床应用和流行病学调查.

Objective To compare the advantage and disadvantage of fluorescent real-time polymerase chain reaction (FQ-PCR) with the conventional tissue culture method, hemagglutination test and serological subtyping influenza A virus. The aim is to establish a convenient and specific method for the identification of influenza A virus from clinical samples. Method 76 influenza virus isolates were used in the testing of the sensitivity and specificity of FQ-PCR. Two-fold serially diluted influenza A (H1N1 and H3N2 subtypes) was used in the testing of the sensitivity of FQ-PCR. The concentration of virus was determined hy hemagglutination test and titration on tissue culture. The sensitivity and specificity of FQ-PCR were also tested on 684 pharyngeal swab samples. Result 60 out of 76 infleunza virus infected cell culture supernatant were positively identified by FQ-PCR. The result was consistent with the conventional tissue culture, hemagglutination test and serological subtyping methods. The sensitivity of FQ-PCR is higher,in an order of 25～26, than the conventional tissue culture method. Conclusion FQ-PCR is a rapid, specific, and sensitive method for the detection of influenza A virus.