Establishment and preliminary evaluation of a non-invasive fetal M blood group genotyping method by real-time PCR
10.13303/j.cjbt.issn.1004-549x.2026.04.011
- VernacularTitle:无创胎儿M血型抗原实时荧光定量PCR基因分型方法的建立及初步评价
- Author:
Shuangshuang JIA
1
;
Chunyan MO
1
;
Ling WEI
1
;
Jizhi WEN
1
;
Runqing ZHANG
1
;
Yanli JI
1
Author Information
1. Guangzhou Blood Center, Guangzhou 510095, China; Institute of Blood Transfusion and Hematology, Guangzhou Medical University, Guangzhou 510095, China; The Key Medical Laboratory of Guangzhou, Guangzhou 510095, China
- Publication Type:Journal Article
- Keywords:
anti-M related hemolytic disease of the fetus and newborn (HDFN);
cell-free fetal DNA (cff-DNA);
non-invasive fetal M antigen genotyping;
real-time fluorescent quantitative PCR
- From:
Chinese Journal of Blood Transfusion
2026;39(4):493-500
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To establish a method for the genotyping of fetal M blood group antigen by extracting cell-free fetal DNA (cff-DNA) from maternal plasma, so as to guide the management of M antigen-negative pregnant women with IgG anti-M antibody during pregnancy. Methods: A realtime fluorescent quantitative PCR (realtime PCR) method was established. The specificity and sensitivity of the method were validated by dilution of genomic DNA. Subsequently, a total of 12 M antigen-negative pregnant women were enrolled. The cff-DNA was extracted from maternal plasma, and fetal M antigen genotyping was performed by realtime PCR. Fetuses were classified as M-positive or M-negative according to the presence or absence of amplification curve. The accuracy of the method was validated by comparing fetal M antigen genotyping results with the serological results using the cord or peripheral blood of the neonate at birth. Results: Among the 12 M antigen-negative pregnant women, anti-M was detected in five cases, of which four cases had IgG anti-M, and one case had fetal anemia. The results of fetal M antigen genotyping showed that 9 cases were M-positive (9/12, 75%) and 3 cases were M-negative (3/12, 25%). Serological results of blood samples collected after birth from four M-positive fetuses and one M-negative fetus were consistent with the genotyping results. Conclusion: We have, for the first time, established a non-invasive prenatal genotyping method for fetal M antigen using maternal plasma cff-DNA, and preliminarily demonstrated the feasibility of this method.
