中国输血杂志2025，Vol.38Issue(3)：368-374,7.DOI:10.13303/j.cjbt.issn.1004-549x.2025.03.010
无创胎儿游离DNA RHE血型基因型诊断技术的准确性和可行性研究
Accuracy and feasibility of non-invasive cell-free fetal DNA RhE blood group genotyping
摘要
Abstract
Objective To explore the accuracy and feasibility of non-invasive prenatal diagnosis of fetal RhE genotype using cell-free fetal DNA(cff-DNA)from maternal peripheral blood.Methods A total of 134 pregnant women with single fetuses and RhE-negative blood group were selected from our hospital from November 2023 to August 2024.Free DNA ex-traction kit was used to extract free DNA from peripheral blood of pregnant women,and the RhE blood group genotype of free DNA was detected by real-time fluorescent quantitative PCR(RT-qPCR).If the qPCR amplification signal of the sam-ple was negative,the methylated RASSF1A gene was amplified,and the positive amplification result was used as a sign of successful extraction of cff-DNA.Serological microcolumn gel method was used to detect the phenotype of RhE blood group in neonatal peripheral blood.Results Among the 134 maternal peripheral blood samples,the cff-DNA detection of RhE blood group phenotypes was consistent with the RhE blood group genotyping of neonatal peripheral blood in 133 cases,inclu-ding 90 cases of Rhee genotype and 43 cases of RhE genotype,with diagnostic concordance rate of 99.3%,sensitivity of 97.7%,specificity of 100%,youden index of 0.977,area under ROC curve of 0.995,the Kappa value of 0.983,positive predictive value of 100%,and negative predictive value of 98.9%.The sample of 1 case failed to be detected.After the amplification of methylated RASSFIA gene,it was confirmed that the reason for the failure was that no cff-DNA was extracted from the sample.The diagnostic concordance rates of the first,second and third trimesters were 93.8%(15/16),100%(51/51)and 100%(67/67),respectively.Fisher′s exact test method was used to calculate the P value,which was P>0.05,indicating that there was no statistical significance in the difference of diagnostic concordance rate among the three pregnancy periods,and there was no difference in the detection concordance rate of this method in different pregnancy peri-ods.Conclusion The use of cff-DNA in maternal peripheral blood for the detection of fetal RhE blood group genotype is an accurate and highly feasible non-invasive prenatal diagnostic method,which is helpful for the clinical diagnosis of fetal and neonatal hemolytic disease caused by anti-E antibody.关键词
无创产前诊断/胎儿RhE血型/胎儿游离DNA/胎儿新生儿溶血病/实时荧光定量PCRKey words
non-invasive prenatal diagnosis/fetal RhE blood group/cell-free fetal DNA(cff-DNA)/hemolytic disease of the fetus and newborn/quantitative real-time PCR分类
医药卫生引用本文复制引用
杨金华,任道菊,李小薇,肖军,游江舟,陈春月,张晓娟,李翠莹..无创胎儿游离DNA RHE血型基因型诊断技术的准确性和可行性研究[J].中国输血杂志,2025,38(3):368-374,7.基金项目
2024年度空军军医大学临床研究计划项目(2024LC2406) （2024LC2406）
