聚合酶链反应检测患儿尿标本中的巨细胞病毒DNA

本文建立了聚合酶链反应(PCR)检测人巨细胞病毒(HCMV)方法,探讨了该方法的某些实验条件。用PCR检测24例临床患儿尿标本。证明PCR方法特异、敏感、简便而快速。同科不同属病毒如HSV、EBV等以及人源正常细胞均无假阳性结果:最小检出量可达0.1fg DNA,相当于6个基因拷贝;用水浴箱手控时间即可进行PCR循环,30次循环仅需97min30s;PCR和DNA杂交检测尿标本中HCMV阳性例数分别为20和14例。该结果证明HCMV感染儿童的广泛性和严重性,初步表明儿童肾病综合症与HCMV有关。

Detection of human cytomegalovirus DNA in urine from ill children by polymerase chain reaction

The polymerase chain reaction ( PCR ) assay for the detection of human cyto-megalovirus ( HCMV ) was developed in our laboratory. Some reaction conditions were found out and 24 urine specimens were detected by PCR. The PCR assay was specific,sensitive, simple and rapid. It can be performed by using three bathes and controlling time by stop-watch. No cross-reactivity to Herpesvirus, EB virus or normal human embryo lung cell was observed, only 0.1fg of HCMV-AD169 strain DNA EcoRI-J-fragment can be detected after 30 cycles of amplification, corresponding to approximately 6 gene copies. The numbers of positive specimens of PCR and DNA hybridization for detecting HCMV in urine from the children with hepatitis or nephropathy syndrome were 14 and 20 respectively. This shows HCMV infection is extensive and serious, and nephropathy syndrome may be related with HCMV.