采用DNA-DNA杂交试验检测人巨细胞病毒感染的初步研究

DETECTION OF HUMAN CYTOMEGALOVIRUS INFECTION BY DNA-DNA HYBRIDIZATION

建立了快速的DNA-DNA杂交试验检测感染人巨细胞病毒(HCMV)的细胞.采用32P或生物素标记的克隆的DNAJ片段作为探针.实验结果表明该探针是敏感的、特异的,能检测出0.9pg(32P-探针)或90pg(生物素-探针)的同源DNA.该探针不能和其它疱疹病毒、未感染人胚肺细胞和人胚DNA相杂交.本试验检测了先天性CMV感染的新生儿、孕妇和助血员的尿标本144份和血标本154份,获得了满意的结果.

A rapid DNA-DNA hybridization has been established to identify the human cytomegalovirus (HCMV) in infected cells by cloned viral DNA J fragment labeled with 32P or Biotin as a probe.Preliminary experiments showed that the probe is sensitive and specific to detect 0.9Pg(32P-probe) or 90pg(Biotin-probe) homologous DNA,60 cells(32P-probe)or 325 cells(Biotin-probe) of HCMV infected fibroblasts.The labeled probe fails to hybridize with DNA from other herpes viruse,uninfected human lung fibroblasts and placental DNA.One hundred forty-four urine specimens and 154 White blood cell specimens collected from the new born infants with congenital CMV infection,pregnant women and blood donors have been detected for HCMV by DNA-DNA hybridization with satisfactory results.