摘要
采用缺口转移(Nick-translation)方法标记克隆化的乙型肝炎病毒(HBV)DNA制备探针,能特异地与HBV DNA杂交,在本实验条件下,其最低检测量为0.2pg,同位素参入率为23.5—67.0％,使用100μCi ^(32)P dCTP,比放射活性为10~8cpm//μg DNA左右。临床血清检测统计分析表明:HBV DNA与HBsAg、HBeAg、HBcAg、DNAP的阳性符合率分别为53.3％、69.7％、78.6％、81.7％,其灵敏性和特异性均比免疫学检测法高。
Hepatitis B virus(HBV) DNA purified from the cloned plasmid pHBV_1 and labelled by nick-translation with α-^(32)p dCTP was as probe, which can be hybridized specifically with HBV DNA. The minimum amount about 0.2pg of DNA has been detected by high specific activity probes. The range of the isotope incorporation rates were 23.5-67.0 per cent. The probes's specific activities were 1-1.50×10~8cpm/μg DNA. According to clinical statistics the positive proportions of HBV DNA Correspond to 53.3% 69.7% 78.6% and 81.7% for HBsAg, HBeAg and HBcAg as well as DNAp respectivity The HBV DNA hybdization method for serum detection is more sensitive and specific than immunoassay methods.
关键词
乙肝病毒
DNA探针
血清检测
Hepatitis B virus
Probe
Nick translation
Serum detection
