免疫磁珠法在预测胎儿RHD血型中的应用  

Application of magnetic beads method in predicting fetal RHD blood group

作　　者：王娇 郭珈[1] 葛昕[1] 付有荣[1] 沈长新[1] 

机构地区：[1]武汉大学中南医院输血科,武汉430071

出　　处：《临床血液学杂志（输血与检验）》2018年第1期108-112,共5页Journal of Clinical Hematology(Blood Transfusion & Laboratory Medicine)

基　　金：湖北省卫生计生委采供血专项课题(No:WJ2015CB005)

摘　　要：目的:探索应用免疫磁珠法富集母体外周血DNA并预测其胎儿RHD血型。方法:采集96例来自武汉大学中南医院产前诊断确诊为RHD阴性孕妇外周血,使用免疫磁珠法和血液DNA提取试剂盒(QIAgene DNA法)抽提母体外周血DNA用以提取胎儿DNA。通过测定母体血浆中的SRY和STR来证实是否获得胎儿DNA,同时使用荧光定量PCR(RQ-PCR)扩增胎儿RHD基因外显子7,预测胎儿RHD血型。结果:通过免疫磁珠法,胎儿DNA的总提取量提高了1.8倍。配对四格表2检验结果显示免疫磁珠法在准确度方面与QIAgene DNA法相较,妊娠早期(≤14周)2种方法差异有统计学意义(P<0.05),而妊娠中晚期(>14周)2种方法差异无统计学意义。另外,使用免疫磁珠法提取DNA预测胎儿RHD血型时,有86例结果与胎儿RHD血清学方法鉴定一致,使胎儿RHD血型预测的准确率高达89.6%。结论:使用免疫磁珠法提取胎儿DNA不仅能够改善胎儿DNA的总提取量,同时还可以提高妊娠早期胎儿RHD血型预测的准确率,为RHD血型不合导致的新生儿溶血病的早期诊断和治疗提供更好的试验基础。Objective：To explore enrich maternal peripheral blood DNA using the magnetic beads method in the application of predicting fetal RHD blood group.Method：Peripheral blood of 96 RHD negative pregnant women was collected from Zhongnan Hospital of Wuhan University.Magnetic beads method and blood DNA extraction kit（QIAgene DNA method）enriching the maternal peripheral blood DNA were used to extract fetal DNA.Fetal DNA was confirmed by testing SRY and short tandem repeat（STR）in the maternal plasma.In the meanwhile,exon 7 in the RHD gene was amplified using RQ-PCR to which could be applied to predict the fetal RHD blood group.Result：Using the magnetic beads method,the overall yield of fetal DNA was improved by a mean factor of1.8.The results indicated that there was a significant difference in accuracy of fetal RHD blood group between the two methods（P〈0.05）in the early gestation（less than 14 weeks）,while in the middle and late gestation（exceeded 14 weeks）there was not a significant difference in accuracy of fetal RHD blood group between the two methods.In addition,when using magnetic beads method to extract DNA was used to predict fetal RHD blood group,the results of 86 cases were in accordance with fetal serum RHD identification method,which made its accuracy of fetal RHD blood group up to 89.6%.Conclusion：Using the magnetic beads method could not only improve the overall yield of fetal DNA,but also enhance the accuracy of fetal RHD blood group in the early gestation（less than14 weeks）.Then it will provide a better experimental basis for the early diagnosis and treatment of neonatal hemolytic disease caused by incompatibility of RHD blood group.

关 键 词：免疫磁珠法 母体外周血DNA RHD血型 

分 类 号：R457.1[医药卫生—治疗学]