荧光定量PCR法在不同孕期通过母体血浆检测胎儿RhCcE血型

Determination of the fetal RhC,c,and E genotypes from maternal plasma in different gestational ages by FQ-PCR

目的比较荧光定量PCR法(FQ-PCR)在不同孕期通过母体血浆检测胎儿RhCcE血型的准确性。方法随机采集孕期8～38周的孕妇外周血334份,分离血浆,提取游离胎儿DNA,用FQ-PCR方法检测胎儿RhCcE血型,利用胎儿脐带血标本检测的结果做验证。结果在妊娠8～≤12、13～≤20及21～38周的孕妇中,RhCcE血型各基因型的FQ-PCR检测结果与脐带血血清学检测结果比较:C基因型正确率分别为94.1%(16/17)、91.6%(22/24)、100%(18/18),漏检率为5.9%(1/17)、8.3%(2/24)、0%(0/18);c基因型正确率分别为97.6%(41/42)、100%(55/55)、100%(30/30),漏检率为2.4%(1/42)、0%(0/55)、0%(0/30);E基因型正确率分别为97.8%(46/47)、98.4%(60/61)、100%(35/35),漏检率为2.1%(1/47)、1.6%(1/61)、0%(0/35)。结论随着孕期的增加,FQ-PCR法检测胎儿RhCcE血型的准确性相应增加,孕晚期时达100%。

Objective To study the accuracy of fluorescence quantitative PCR（FQ-PCR） assay in the detection of fetal RhC,c,and E genotypes from maternal plasma at different gestational ages.Methods Free fetal DNA were extracted from 334 plasma samples of pregnant women at 8~38 weeks of gestation.The fetal RhC,c,E genotypes were identified by FQ-PCR assay.The RhC,c,E phenotype of umbilical cord blood from infant was tested by serologic method.Results The genotype results of FQ-PCR assay from maternal plasma at gestational weeks of 8~≤12 weeks,13~≤20 weeks,and 21~38 weeks were compared to the phenotype results of umbilical cord blood of infants.The accuracy rates of C genotyping were 94.1%（16/17）,91.6%（22/24）,and 100%（18/18）,respectively,with respective missing rates of 5.9%（1/17）,8.3%（2/24）,and 0%（0/18）.The accuracy rates of c genotyping were 97.6%（41/42）,100%（55/55）,and 100%（30/30）,respectively,with respective missing rates of 2.4%（1/42）,0%（0/55）,and 0%（0/30）.The accuracy of E genotyping was 97.8%（46/47）,98.4%（60/61）,and 100%（35/35）,respectively,with respective missing rates of 2.1%（1/47）,1.6%（1/61）,and 0%（0/35）.Conclusion With increasing of gestation age,the accuracy of fetal RhC,c,and E genotyping by FQ-PCR will reach up to 100% in the third trimester of pregnancy.