非放射性乙型肝炎病毒核酸探针的制备和应用

Preparation and Application of Non-radiolabeled Hepatitis B Virus Nucleic Acid Probe

非放射性的生物素（Ｂｉｏｔｉｎ）和地高辛（Ｄｉｇｏｘｉｇｅｎｉｎ）标记的乙肝病原基因探针（Ｂｉｏ－ＨＢＶＤＮＡ和Ｄｉｇ－ＨＢＶＤＮＡ探针），特异性强，灵敏度高，最低检出值分别为１ｐｇ和０．５ｐｇ。３２０份血清检测了ＨＢＶＤＮＡ和ＨＢｅＡｇ，并与α－_（３２）Ｐ标记的探针检测结果相比较，它们的特异性分别为９６．５％、９８．１％和８８．９％；灵敏度分别为８１．７％、８６．７％和６１．７％；符合率则为９３．８％、９５．９％和８３．８％。应用Ｄｉｇ－ＨＢＶＤＮＡ探针，作斑点杂交试验，在乙肝患者血清、白细胞、尿、粪、唾液以及旅客列车的桌面、车箱门把手、厕所门把手和水笼头上，均可检得阳性结果。我所研制的非放射性乙型肝炎病原基因诊断试剂，为科学研究和临床检测提供了极为有用的工具。

Non-radiolabeled HBV DNA probes (biotin-HBV DNA and digoxigenin-HBV DNA probes) of high specificity and sensitivity were prepared with a minimal detection limit of 1pg and 0.5pg,respectively. HBV DNA and HBeAg were detected in 320 serum samples,and the results were compared with those obtained by α-32P-HBV DNA probe. The specificity of both Bio-HBV DNA and Dig-HBV DNA probes and HBeAg test was 96.5%,98.1% and 88.9%,the sensitivity 81.7%, 86.7% and 61.7% and the consistency 93.8%, 95.9% and 83.8%,respectively. The positivity for HBV DNA was found in sera, white blood cells, urine, feces,saliva from patients with hepatitis B and enviromental specimens (table surface,handles and faucets of railway carriages)by dot hybridization with Dig-HBV DNA.