Journal of Reproduction & Infertility

Journal of Reproduction & Infertility

Noninvasive Prenatal Diagnosis of Fetal RHD Status Using Cell-free Fetal DNA in Maternal Plasma

Authors
Mohammad Hossein Ahmadi 1
Ali Akbar Pourfathollah 2
Maryam Rabiee 3
Naser Amirizadeh 4
1 Department of Laboratory Sciences, School of Paramedical Sciences, Mashhad University of Medical Sciences, Mashhad, Iran; Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran
2 Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
3 Department of Obstetrics and Gynecology, Shahed University, Tehran, Iran
4 Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran
10.18502/jri.v23i2.8998
Abstract
Background: The main cause of hemolytic disease of the fetus and newborn (HDFN) is the incompatibility of the RHD antigen between mother and fetus. Following the discovery of cell-free fetal DNA (cffDNA), noninvasive fetal RHD genotyping also became possible, which will help in the better management of immunized RHD negative mothers and in the targeted prenatal injection of Rho(D) immune globulin (RhIG). The objective of this study was to establish a reliable method with high accuracy to determine the fetal RHD genotype. Methods: The project was a prospective observational cohort study. After cell-free DNA (cfDNA) extraction from maternal plasma, fetal RHD genotyping was performed by duplex real-time polymerase chain reaction (PCR) and exons 5, 7, and 10 of the RHD gene were examined. SRY and RASSF1A genes were used as internal controls to confirm the presence of cffDNA in maternal plasma. Results: Out of 40 samples, 33 were RhD positive heterozygous mothers and 7 cases were RHD negative. In three cases where both the fetal RHD and SRY genotypes were negative, RASSF1A was amplified in cell-free DNA sample treated with the BstUI enzyme, and the presence of cffDNA was confirmed. Conclusion: The findings reveal that the strategy used in this study is reliable and it is possible to determine the fetal RHD status with high accuracy. The strategy can help targeted injection of RhIG and prevent unnecessary injection in RhD negative mothers who carry an RhD negative fetus.
Keywords
cell
free fetal DNA
Fetal RHD genotyping
Hemolytic disease of the fetus and newborn
Noninvasive prenatal diagnosis
Rho(D) immune globulin
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Journal of Reproduction & Infertility
Volume 23, Issue 2
Spring 2022
Pages 128-134