地高辛标记HBV DNA探针的应用

Application of HBV DNA probe labelled with digoxigenin

按随机引物法以地高辛标记HBV DNA探针,与按缺口翻译法以^(32)P标记的探针进行了比较。这两种探针与乙肝血源疫苗及患者血清标本中HBV DNA进行杂交,结果两种探针均可要出1pg同源序列DNA,而地高辛标记的探针,在0.1Pg水平仍可呈现弱着色反应。该探针在一20℃储存6个月,其敏感性不减弱。这种探针在常规诊断实验室更易推广使用。

The whole HBV DNA sequences were labelled by random primer incorporation of Digoxigenin - labelled dUTP, and compared with ^(32)P-labelled probe by nick translation. The two probes were used to detect HBV DNA in plasma- derived HB Vaccine and sera of clinical patients by using Dot-blot hybridization. The results showed that the sensitivity of the two probes were comparable.Both of them could detect 1 pg of homologous DNA, but for the 0.1 pg level of HBV DNA could be detected only with a faint coloured precipitate by digoxigen-labelled probe. The stability experiments of the probe demonstrated that the sensitivity did not decreese after 6 months at -20℃. The non-isotope labelled probe is more readily applicable in routine diagnostic laboratories.