人巨细胞病毒的生物素DNA探针的制备和应用

Preparation and Application of Biotin-labelled Human Cytomegalovirus Nucleic Acid Probe

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。

A diagnostic assay has been developed to detect human cytomegalovirus (HCMV)DNA in clinical specimens with biotin-labelled DNA probe, cloned fragment of HCMV strain ADl69.The biotin-labelled probe can detect 100 pg of homologous DNA and does not hybridize with DNA from other herpesvirus or human embryonic lung cells(HEL) and Hep-2 cells. 30 umbilical cord blood and 10 urine specimens from pregnant women were examined by hybridization assay with biotin-labelled probe. Among 30 umbilical cord blood specimens, 11(30%) and, among 10 urine specimens, 3(33%) were positive for HCMV DNA respectively. The results suggest that the DNA-DNA hybridization assay with biotin-labelled DNA probe is high specific and sensitive, and more rapid than routine tissuse culture techniques.Biotinlabelled HCMV probe can be used in clinical diagnosis of human cytomegalovirus infection.