孕妇外周血胎儿细胞中巨细胞病毒基因的检测及意义

Analysis of cytomegalovirus gene by isolating fetal cells in maternal blood

目的 利用孕妇外周血中胎儿细胞进行无创性产前诊断巨细胞病毒宫内感染的新方法。方法 (1)采用显微操作技术从 2 73例孕妇外血中分离单个胎儿有核红细胞。 (2 )应用多重引物原位合成技术 (primedinsitulabeling ,PRINS)检测 76例孕妇外周血HCMV DNA阳性标本的单个胎儿细胞的SRY基因和HCMV DNA基因。 (3)应用引物延伸预扩增法 (primedextensionpreamplification ,PEP)及聚合酶链反应 (PCR)检测 2 73例孕妇外周血中单个胎儿细胞的SRY基因和HCMV DNA基因。结果 (1)应用显微操作技术分选胎儿细胞的分选率为 10 0 %。 (2 )PRINS技术检测SRY基因的敏感性为 97 5 6 % (40 / 4 1) ,特异性为 10 0 % (35 / 35 )。检测HCMV DNA基因的敏感性为 92 6 8% (38/ 4 1) ,特异性为 10 0 % (35 / 35 )。 (3)PEP及PCR方法检测SRY基因的敏感性为97 39% (14 9/ 15 3) ,特异性为 99 17% (119/ 12 0 ) ,检测HCMV DNA基因的敏感性为 95 12 % (39/4 1) ,特异性为 10 0 % (2 32 / 2 32 )。结论 应用PRINS技术、PEP方法及PCR技术对孕妇外周血中单个胎儿细胞进行非创伤性产前诊断巨细胞病毒宫内感染是一种敏感性高特异性强的新方法 ,可能具有广阔的临床应用前景。

Objective To establish a new criteria of noninvasive prenatal diagnosis of human cytomegalovirus intrauterine infection by isolating single fetal cell in maternal peripheral blood Methods (1) Micromanipulation techniques were applied to isolate single fetal nucleated erythroblasts from 273 maternal blood samples (2) SRY gene and HCMV DNA of the single fetal cell were detected by multiple primed in situ labeling (PRINS) from 76 samples of maternal peripheral blood which were HCMV DNA positive (3) SRY gene and HCMV DNA of a single fetal cell were detected by primed extension preamplification (PEP) and polymerase chain reaction (PCR) from 273 samples of maternal peripheral blood Results (1) The detection rate of fetal cells from maternal blood was 100% by micromanipulation techniques (2) The sensitivity of detecting SRY gene by PRINS was 97 56% (40/41) and its specificity was 100%(35/35) (3) The sensitivity and specificity of detecting SRY gene by PEP and PCR were 97 39% (149/153)and 99 17%(119/120)respectively The sensitivity and specificity of detecting HCMV DNA by PEP and PCR were 95 12%(39/41)and 100%(232/232)respectively Conclusions The new method of noninvasive prenatal diagnosis of human cytomegalovirus intrauterine infection by isolating single fetal cell in maternal peripheral blood by use of PRINS, PEP and PCR was of high sensitivity and specificity and may be widely used in clinical laboratory