荧光定量PCR法从母体血浆检测胎儿RhCcE血型

FQ-PCR testing for the determination of the fetal RhCcE factor in maternal plasma

目的建立荧光定量PCR(FQ-PCR)法从母体血浆中检测胎儿RhCcE血型,用于胎儿和新生儿溶血病的产前诊断。方法采集孕期8～20周的妇女血浆,采用QIAampDNA Mini方法从母体外周血中提取游离胎儿DNA,用FQ-PCR法检测胎儿DNA的RhCcE基因型,预测胎儿血型。与胎儿脐带血标本检测的结果进行比较,评价采用FQ-PCR法从母体检测胎儿RhCcE血型的准确性。结果 46例RhE阴性母体血浆中检测出18例RhE血型阳性,与胎儿脐带血检测结果完全一致,准确率100%;36例Rhc阴性母体血浆中检测出24例Rhc血型阳性,与胎儿脐带血检测结果完全一致,准确率100%;从35例RhC阴性母体血浆中检测出10例RhC血型阳性,与胎儿脐带血检测结果相比漏检2例,准确率94%。结论 FQ-PCR法能够在孕早期检测出胎儿Rh-CcE血型,可以用于胎儿和新生儿溶血病的产前诊断。

Objective To establish a fluorescent quantitative PCR（FQ-PCR） method to detect fetal RhCcE factor in maternal plasma,which contribute to fetal and newborn hemolytic disease prenatal diagnosis.Methods Free fetal DNA were extracted from pregnant women at 8～20 week of gestation.FQ-PCR method was used to detect the fetal DNA RhCcE genotype to predict the fetal blood group,compared with results of serological testing from umbilical cord blood of infant.Results 18 cases were positive in RhE genotyping of FQ-PCR from 46 RhE negative maternal,the same results as serological testing from umbilical cord blood of infant.The accuracy rate was 100%;24 cases were positive in Rhc genotyping from 36 Rhc negative maternal,the same results with serological testing from umbilical cord blood of infant.The accuracy rate was 100%;10 cases were positive in RhC genotyping from 35 RhC negative maternal,but 12 cases were positive in serological testing from umbilical cord blood of infant.The accuracy rate was 94%.Conclusion FQ-PCR can be used to detect fetal RhCcE factor in early gestation,prenatal diagnosis of fetal and newborn hemolytic disease.