摘要
在人巨细胞病毒(HCMV)高度保守区IEA基因内设计一对引物,建立了聚合酶链反应(PCR)检测HCMV DNA的方法,经实验证明有高度的特异性和敏感性,对其它疱疹病毒和正常人DNA无交叉反应,HCMV AD169株DNA EcoRI酶切J片段的最小检出量为0.1fg,相当于6个基因拷贝。对PCR反应体系中的有关实验条件如Mg^(2+)浓度、引物浓度、Taq DNA多聚酶的浓度、循环程序等及尿标本的处理方法进行优化选择。用PGR检测20例肾移植受者尿标本中HCMV DNA,结果11例(55％)阳性。表明PGR是一种快速、特异、敏感的检测方法,适用于肾移植等病人的HGMV感染监测。
Polymerase chain reaction (PCR) for detecting humam cytomegalovirus (HCMV) has been established and its specificity and sensitivity ascertained. There was no cross-reaction with herpesvirus or humam DNA. The lowest detection level was 0.1fg of HCMV-AD 169 strain DNA EcoRI-J-fragment, corres ponding to approximately 6 gene copies. The experimental conditions were optimized including Mg2+ concentrations, primer concentrations, TaqDNA polymerase concentrations, temperature cycling protocols and methods of DNA extraction. Urine specimens of 20 renal transplant recipients were examined with PCR for HCMV and the positive rate was 55%. PCR was believed to be a rapid, sensitive and specific technique for the detectiom of HCMV infection.
出处
《空军总医院学报》
1993年第2期67-69,共3页
Journal of General Hospital of Air Force,PLA
