简化HBV-DNA抽提用于多聚酶链式基因扩增

A SIMPLIFIED TECHNIQUE OF HBVDNA ISOLATION IN PCR

采用异硫氰酸胍、柠檬酸钠、2-巯基乙醇、十二烷基肌氨酸钠作为变性裂解剂,结合水饱和酚、氯仿、异戍醇等进行快速程序式抽提血清HBV-DNA模板用于多聚酶链式(PCR)基因扩增。该提取法比一般方法简单、稳定、不易污染,抽提过程可在lh内完成。PCR应用结果表明敏感性高于地高辛素标记探针斑点杂交法。经酶切及a-^(32)标记放射自显影鉴定PCR产物具有良好的特异性。

A new method of HBV-DNA isolation by a simple extraction with an acid guanidi-nium thiocyanate-phenol saturated with water in PCR was described. The method was simple and can be completed in 1 hour. Compared with spot hybridizition using probe labelled by digoxi-genin for detection of HBV-DNA, this technique had a high sensitivity, and was particular useful in processing large number of clinical samples.