HBV套式PCR技术的建立及HBcAb血清的检测

The Establishment of a Nest-PCR Technique and the Detection of HBcAb Serums for HBV DNA

本文实验设计了套式ＰＣＲ引物进行ＨＢＶＤＮＡ诊断。外引物限定ＨＢＶＣ基因的一个６１３ｂｐ片段，内引物限定其内－５０７ｂｐ的片段，扩增后产物被限制性内切酶图谱证明。该技术的特异性强，重复性好，灵敏性达到１－１０ｆｇ，对ＨＢＶ血清可做出明确诊断。应用这项技术，对４２例ＨＢｃＡｂ血清进行了检测，实验表明仅ＨＢｃＡｂ阳性血清同带ＨＢｓＡｇ的ＨＢｃＡｂ阳性血清一样，体内持续进行着大量ＨＢＶＤＡＮ复制。

A nest-PCR technique was developed for HBV DNA in this study. PCR primers resulted in the expected segments from HBV C gene. Those were demonstrated by the restriction enzymes. The detection limit of the technique for the cloned HBV DNA was 1-10 fg. The technique could be used in clinical diagnosis because of better specificity and repeat. The diagnosis regents for HBV DNA was developed by the technique.42 samples of HBcAb serum were detected for HBV DNA in the assay. Among them 10 f12 serum samples with HBsAg mark,4 of 13 serum samples with HBsAb mark and 15 of 17 serum samples only with HBcAb mark were found to have HBV DNA. The result showed that there was a great deal of sustained HBV DNA duplication in body of the patients only with HBcAb mark as in the ones with HBsAg mark.