PCR结合地高辛标记探针杂交检验乙型肝炎病毒C基因启动子

应用聚合酶链式反应结合地高辛标记探针杂交方法检测乙型肝炎病毒C基因启动(HBV,CP)。对聚合酶链反应(PCR)法扩增的HBV DNA直接测序进行DNA同源性分析。并运用探针杂交对结果进行进一步鉴定;应用PCR法扩增20份患者血清,阳性率达95％(19/20),同时运用探针杂交进行进一步检验,与PCR结果符合率达90％以上,并选择中度、重度乙肝患者血清和pGEM,7Z—HBV质粒扩增的HBV.CP各一份分别进行测序,与报告基因的同源性分别为72％、66.5％、90％。建立了特异敏感的检测乙型肝炎病毒C基因启动子(HBV.CP)的PCR方法,可用于HBV基因变异规律的研究。同时初步的研究结果表明,肝炎病人的HBV.CP区存在较多的变异,可能与病情轻重有一定的相关性。

DETECT HEPATITIS B VIRUS CORE PROMOTER IN PATIENT BY USING PCR LIGATED WITH DIGOXIGENIN LABELED PROBE HYDRIZATION

To explpore effective method to detect HBV. CP in hepatitis patients. Polymerase chain reaction amplified HBV DNA fiagments were directly sequenced and analyzed with the help of computer. On the other hand, digoxigenin labeled probe also be used in order to determine PCR results. With the help of PCR, the positive rate of serum samples from 20 HB patients were 95% (19/20). The rate of correspondence of PCI? and probe methods can reach more than 90%. The HBV. CP fragments of two patients and pGEM. 7Z -HBV plasmid were sequenced. Compared with repor gene sequences, the identical rates of 2 patients were only 72% and 66. 5%, but pGEM 7Z -HBV plasmid was 90%. The PCR above is proved to be a sensitive and specific method for HBV detection and the differentiation of HBV. CP gene of patients was largely. The CP mutations in patients may relate to the state of an ill-