Accuracy and feasibility of non-invasive cell-free fetal DNA RhE blood group genotyping
10.13303/j.cjbt.issn.1004-549x.2025.03.010
- VernacularTitle: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
- Author:
Jinhua YANG
1
;
Daoju REN
2
;
Xiaowei LI
3
;
Jun XIAO
3
;
Jiangzhou YOU
3
;
Chunyue CHEN
3
;
Xiaojuan ZHANG
3
;
Cuiying LI
1
,
4
Author Information
1. Graduate School of PLA Medical College, Beijing 100853, China
2. Air Force Clinical College, The Fifth Clinical Medical College, Anhui Medical University, Hefei 230032, China
3. Department of Blood Transfusion, Air Force Specialty Medical Center, Air Force Medical University, Beijing 100142, China
4. Department of Blood Transfusion, Air Force Specialty Medical Center, Air Force Medical University, Beijing 100142, China
- Publication Type:Journal Article
- Keywords:
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
- From:
Chinese Journal of Blood Transfusion
2025;38(3):368-374
- CountryChina
- Language:Chinese
-
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 extraction 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 sample 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, including 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 periods. [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.
