运用PCR-SSP技术对羊水细胞ABO血型基因定型研究  

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

作　　者：袁晖[1] 林春燕[1] 王晨虹[1] 谢建生[1] 

机构地区：[1]深圳市妇幼保健院,518028

出　　处：《中国优生与遗传杂志》2011年第5期12-14,共3页Chinese Journal of Birth Health & Heredity

基　　金：深圳市科技和信息局资助项目(项目编号:200803075)

摘　　要：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。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 the two methods.Took 1.0~2.0ml venous blood from 40 babies have been born with and was detected ABO blood group by general method and PCR-SSP.And Comparison with the ABO blood group of fetal and baby.Results： Among 80 fetal there were 79 successfully detected by PCR-SSP and DHPLC techniques,21 with Blood type A,19 with blood type B and 38 with blood type O.The genotyping of ABO blood type detected by PCR-SSP was coincidence with the detecting by DHPLC.Genotyping of ABO blood group by PCR-SSP in 40 infant was same as the results of the general method testing,and the blood group of amniotic fluid cells of prenatal diagnosis was identical to the diagnosis after delivery.Conclusion： Testing fetal ABO blood group by PCR-SSP with amniotic fluid cell was accurate and reliable prenatal diagnosing method.And provide the basis for clinical use of the therapy on pregnant woman who was significantly high antibody to A（B）.

关 键 词：ABO血型基因型 ABO母儿血型不合 羊水细胞 产前诊断 PCR-SSP技术 

分 类 号：R440[医药卫生—诊断学]