生物素探针斑点杂交法检测血清HBV-DNA

DOT-BLOT HYBRIDIZATION ASSAY FOR THE DETECTION OF HEPATITIS B VIRUS DNA IN SERUM

用生物素代替^(32)P自行标记HBV-DNA探针及自己配制的试剂检测28例乙肝患者血清HBV-DNA,结果19例血清存在HBV-DNA,占67％。其中9例HBeAg阳性患者,HBV-DNA均为阳性。我们认为此法敏感性高,重复性好,简便,而且可以半定量。因此可以在临床上推广应用。

The detection of hepatitis B virus (HBV) DNA in patient serum has been shown to be one of the most important parameters for the diagnosis of hepatitis B, and for the evaluation of pathological changes of liver and prognosis in patients. In the past, this nucleic acid hybridization technique was mostly based on the ^(32)P-labeling HBV-DNA probes. Because of disadvantages of radioactive isotopes, the radiolabeled probe based assay system was not suitabe for routine clinical laboratories. To overcome such limitations, we have used non-radioactive substance biotin instead of ^(32)p to label the probe for HBV-DNA. And all other necessary reagents for detection of this virus-DNA were prepared in our laboratory. When biotinylated probes were used to assay HBVDNA in serum by dot-blot hybridization, 19 out of 28 hepatitis B patients were found to be positive. Of the 9 HBeAg positive patients, all were found to be positive. This biotin-avidin system is a quite sensitive, reproducible, relatively simple method. In addition, this method can be used for semiquantitative determination of HBV-DNA and could be widely used for routine clinical laboratory.