运用PCR-SSP技术对羊水细胞ABO血型基因定型研究

目的探讨PCR序列特异性引物（PCR-sequence specific primer,PCR-SSP）技术对胎儿羊水细胞ABO血型基因定型研究,分析母儿血型吻合情况。方法选择2008年1月～2010年2月于我院优生优育门诊及产前诊断中心就诊O型血孕妇80例,其丈夫血型分别为A型40例,B型37例,AB型3例。以上80例孕妇于孕16～21周在B超直视下经腹羊膜腔穿刺抽取羊水20～30ml,其中10ml经高速离心取沉淀的羊水细胞,用常规酚氯仿法提取羊水DNA,运用PCR-SSP技术通过PCR反应特异性扩增ABO基因片段,凝胶电泳检测PCR产物,根据产生的特异性PCR产物来鉴定相应的基因来分析血型。同期运用变性高效液相色谱分析（denaturing high performance liquid chromatography,DHPLC）技术对羊水血型检测结果进行验证。余20ml羊水行细胞培养细胞遗传学分析。另回访40例已出生的胎儿,采集外周静脉血分别运用PCR-SSP技术和常规血清学检测方法鉴定ABO血型,比较产前和产后血型检测结果。结果运用PCR-SSP技术从80例羊水DNA中成功检测79例（79/80,98.75%）,其血型分别为A型21例,B型20例,O型38例。同期运用DHPLC技术对PCR-SSP技术检测结果进行验证,两种检测结果相符。已出生40例婴儿外周血血型分析结果与产前诊断血型结果相符。结论 PCR-SSP技术产前诊断胎儿羊水ABO血型具有可行性,进而分析母儿血型吻合情况,为ABO抗体效价显著增高的孕妇临床用药指导提供实验室依据。

Detecting the Genotyping of ABO blood group of amniotic fluid cell by PCR-SSP

Objective: Detecting amniotic fluid cell by PCR-SSP to accurately diagnose fetal ABO blood group.Methods: 80 pregnant women with O blood group and amniocentesis were carried out at 16 ~21w of gestation.Among their husbands there were 40 men with A blood group,37 with B blood group and 3 with AB blood group.The DNA was extracted from amniotic fluid cells by routing phenol-chloroform method and the ABO blood group was identified directly by PCR-SSP and DHPLC techniques.And the results were compared between th...