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本研究用克隆的HCMV AD169株DNA片段,制备了生物素标记的DNA探针,建立了检测临床脐带血、尿标本中HCMV DNA的核酸探针杂交方法。该探针可测出100pg同源DNA,不与人胚肺细胞、Hep-2细胞DNA以及其他疱疹病毒的DNA发生反应。用核酸杂交方法检测了30份脐带血标本,有11例阳性,阳性率为33%。10例孕妇尿标本中,3例阳性,阳性率为30％。检测结果表明:我们建立的生物素标记的HCMV DNA探针的点杂交法,具有高度的特异性、敏感性,比分离病毒法更迅速,可用于HCMV感染的临床标本的病毒核酸检测。
In this study, a cloned DNA fragment of HCMV AD169 strain was used to prepare a biotin-labeled DNA probe and a nucleic acid probe hybridization method was established for detecting HCMV DNA in clinical umbilical cord blood and urine samples. This probe detects 100 pg of homologous DNA and does not react with DNA from human embryonic lung cells, Hep-2 cells and other herpes viruses. 30 samples of umbilical cord blood were detected by nucleic acid hybridization method, 11 were positive and the positive rate was 33%. Ten cases of urine samples of pregnant women, 3 cases were positive, the positive rate was 30%. The results showed that the spot hybridization method of the biotin-labeled HCMV DNA probe was highly specific and sensitive. It was more rapid than the virus isolation method and could be used for the detection of viral nucleic acids in HCMV-infected clinical samples.