聚合酶链反应结合生物素探针杂交检测HBV DNA的应用

THE COMBINATION OF POLYMERASE CHAIN REACTION AND DNA HYBRIDIZATION ASSAY FOR DETECTION OF SERUM HEPATITIS B VIRUS DNA

本研究将聚合酶链反应和生物素探针点杂交相结合,可直接从5μl 血清中检测到10fgHBV DNA。应用此方法检测20例 HBsAg 和 HBeAg 阳性病例,HBVDNA 均阳性;20例 HBsAg 阳性、HBeAg 阴性病例中,有8例阳性;20例健康献血员中,亦有1例检出 HBV DNA。结果表明这是一种灵敏可靠,适合于基础及临床广泛使用的 HBV DNA 非同位素检测法。

A polyrnerasc chain reaction combined with dot hybridization assay for the detection of hepatitis B virus DNA was developed.Serum HBV DNA was amplied using oligonuc- leotide primers specific for S gene.HBV DVA was determined and confirmed by gel elect- rophoresis and dot hybridization with biotin-labelled HBV probe.HBV DNA could be de- tected in the serum using aliquots as 5 microliters,and the detection limit for cloned he- patitis B DNA was 10 fg.Serum specimens of 20 HBsAg-negative healthy donors and 40 HBsAg-positive chronic hepatitis B patients were assayed.HBV DNA were positive in 1 of the 20 healthy persons,all of the 20 cases of HBeAg-positive patients and 8 of the 20 cases of HBeAg-negative patients by using this PCR assay.The result indicates that-the PCR is much more sensitive than those of serological and dot hybridization methods.