孕妇外周血中胎儿γ血红蛋白基因的检测

Detection of fetal γ globin gene from maternal peripheral blood

目的 探索检测孕妇外周血中胎儿γ血红蛋白基因表达的方法。方法 提取孕妇全血mRNA ,经体外逆转录后 ,用反转录聚合酶链反应 (RT PCR)方法 ,应用ε/γ特异引物引导扩增胎儿ε/γ血红蛋白基因 ,以NdeⅡ酶切后 ,经琼脂糖凝胶电泳 ,在凝胶成像系统下观察。结果 成功自孕妇外周血中扩增出胎儿ε/γ血红蛋白基因 ,ε/γ基因被NdeⅡ酶切成三个γ基因片断 ,大小分别为 191、5 7和 2 6bp。结论 孕妇全血中微量的胎儿mRNA已基本满足体外逆转录及扩增的要求 ,胎儿γ血红蛋白基因的检测可为无创伤性产前诊断的发展奠定基础。

Objective The purpose is to establish a convenient,sensitive and special method as a basis of screening prenatal diseases in population and lay a basis for family plan and clinical application.Methods Blood samples were collected and the fetal mRNA extracted from the pregnant women with the use of random primer.Reverse transcription of mRNA into cDNA was carried out and cDNA was amplified by PCR with the special ε/γ primer used.The product of PCR was cut by enzyme NdeⅡ.All the products underwent electrophoresis in agarose gel.Using 'Gel Works System' to scan the electrophoresis image to detect ε/γ gene band and γ gene band.Results Using RT PCR and agarose gel electrophoresis method,ε/γ gene from peripheral blood of pregnant women was detected successfully,which was cut into three γ gene by NdeII enzyme,the 191 bp,57 bp and 26 bp.Conclusion Using RT PCR to detect fetal ε/γ gene and γ gene from maternal peripheral blood is sensitive and easily accepted by pregnant women. It will be possible to use this method widely in prenatal diagnosis.