聚合酶链反应凝胶呈像技术检测乙型肝炎病毒

目的建立聚合酶链反应（ＰＣＲ） 凝胶呈像（ｇｅｌｄｏｃｕｍｅｎｔａｔｉｏｎｓｙｓｔｅｍｓ,ＧＤＳ）技术,并应用于临床检测乙型肝炎病毒（ＨＢＶ）核酸ＤＮＡ。方法用凝胶呈像技术对血清ＨＢＶ ＤＮＡＰＣＲ扩增产物的电泳凝胶进行摄像、分析,并与紫外灯下的肉眼观察结果相比较,对肉眼未判断出阳性而凝胶呈像分析出阳性的血清标本,用定量ＰＣＲ（ＱＰＣＲ）方法检测ＨＢＶ ＤＮＡ。结果通过３８份血清标本的检测,发现用凝胶呈像技术判断的结果比单纯肉眼观察的阳性率高。前者１９份阳性（５０％）,后者１６份阳性（４２．１％）,符合率为９２．１％。经χ２检验（χ２＝０．４７７,Ｐ＞０．０５）,说明两种方法有很好的一致性。凝胶呈像分析为阳性,而肉眼观察阴性的３份血清标本,经定量ＰＣＲ检测为阳性。结论应用凝胶呈像技术比ＰＣＲ产物电泳后紫外灯下肉眼观察的灵敏度高、客观性强,能避免与紫外线接触,并可长期保留检测结果。

Application of gel documentation systems in HBV detection with PCR

Objective To establish polymerase chain reactiongel documentation system (PCRGDS) technology for detecting deoxyribonucleic acid of hepatities B virus (HBVDNA) in sera. Methods The electrophoresis gel with HBVPCR amplification products was analysed with GDS and observed with naked eyes in ultraviolet. Quantity PCR(QPCR) was employed in the sera whose PCR product was negative in naked eyes observation but positive in GDS analysis. Results In 38 sera samples of patients, the positive rate by GDS was higher than that by naked eyes . The corresponding rate of the two tests was 92.1% (2=0.477, P>0.05). A good coherence was noted between the two methods. Three sera samples positive by GDS and negative by naked eyes were detected positive with QPCR. Conclusion With higher sensitivity and objectivity, GDS avoids the exposure of observers to ultraviolet and preserves the data for a long time.