FOETAL RHESUS D AND SEX GENOTYPING FROM THE PLASMAS OF RH-NEGATIVE PREGNANT WOMEN IN DUHOK

.

  • Sawer Sabri Ahmad Medical Laboratory Technology, College of Health and Medical Technology, Duhok Polytechnic University, Duhok, Iraq
  • SALH ABDULAZEEZ SALH Medical Laboratory Technologist, Duhok Emergency Hospital, Directorate General of Health-Duhok
Keywords: : Rhesus D, , Sex prediction, Fetal DNA, Maternal’s Plasma

Abstract

https://doi.org/10.31386/dmj.2024.18.2.1

 Background. This prospective study was done to evaluate the benefit of Real-time Polymerase Chain Reaction in the determination of RHD and SRY of the fetuses in the plasma of Rhesus D-negative enrolled ladies.

 Methods: Thirty-nine pregnant Rh-negative females with (RhD+) partners were registered in this study. Blood samples were taken from those participants whose pregnancy age was from 20 -28 weeks for the purposes of fetal RHD genotyping and DNA extraction/purification. Real-time PCR was used to determine the RH and sex genotype using particular RHD, SRY, and GLO gene primers and probes.

Results: Out of the total 39 samples, 28/39(71.8 %) were positive for Rhesus D antigen and 11/39 (28.2 %) were negative. The results of prenatal fetal RHD genotyping based on exons 7, and 10 combined were concordant with postnatal RhD phenotyping in 94.9% of cases (37/39) with a sensitivity & specificity of 93.3 and 100% respectively, and precision was 100%. For the gene SYR, there was a concordance rate of 92.3% between the fetal sex genotype and the newborn sex with one false-positive (2.6%) and two false negatives (5.1%) results leading to a precision of 95.2%.

Conclusion: The study demonstrated great accuracy of the Real-Time PCR technique through the usage of cffDNA for the genetic study of the fetal RHD and SYR genes and implicates the effectiveness of this procedure for predicting the necessity of Anti-D immunoprophylaxis in pregnant women whose fetuses have a peril of hemolytic anemia of the newborn due to RhD incompatibility between partners.

Downloads

Download data is not yet available.

References

1. Oliveira J, Osório N, Rocha J, Cruz B, Figueiredo J, Caseiro A, et al. Fetal RHD and RHCE genotyping in plasma of Rh negative pregnant women. Int J Biomed Lab Sci (IJBLS). 2012;1(2):50-8.
2. Daniels G. The molecular genetics of blood group polymorphism. Transplant immunology. 2005;14(3-4):143-53.
3. Avent ND, Reid ME. The Rh blood group system: a review. Blood, The Journal of the American Society of Hematology. 2000;95(2):375-87.
4. Wright CF, Burton H. The use of cell-free fetal nucleic acids in maternal blood for non-invasive prenatal diagnosis. Hum Reprod Update. 2009;15(1):139-51.
5. Van der Schoot CE, Hahn S, Chitty LS, editors. Non-invasive prenatal diagnosis and determination of fetal Rh status. Seminars in Fetal and Neonatal Medicine; 2008: Elsevier.
6. Amaral DR, Credidio DC, Pellegrino J, Jr., Castilho L. Fetal RHD genotyping by analysis of maternal plasma in a mixed population. J Clin Lab Anal. 2011;25(2):100-4.
7. Banch Clausen F. Integration of noninvasive prenatal prediction of fetal blood group into clinical prenatal care. Prenatal Diagnosis. 2014;34(5):409-15.
8. Hromadnikova I, Vechetova L, Vesela K, Benesova B, Doucha J, Vlk R. Non-invasive fetal RHD and RHCE genotyping using real-time PCR testing of maternal plasma in RhD-negative pregnancies. Journal of Histochemistry & Cytochemistry. 2005;53(3):301-5.
9. Lo YD, Tein MS, Lau TK, Haines CJ, Leung TN, Poon PM, et al. Quantitative analysis of fetal DNA in maternal plasma and serum: implications for noninvasive prenatal diagnosis. The American Journal of Human Genetics. 1998;62(4):768-75.
10. Avent ND. The Rhesus blood group system: insights from recent advances in molecular biology. Transfusion medicine reviews. 1999;13(4):245-66.
11. Randen I, Hauge R, Kjeldsen‐Kragh J, Fagerhol M. Prenatal genotyping of RHD and SRY using maternal blood. Vox sanguinis. 2003;85(4):300-6.
12. Schmidt LC, Cabral AC, Faria MA, Monken F, Tarazona-Santos E, Martins ML. Noninvasive fetal RHD genotyping from maternal plasma in an admixed Brazilian population. Genet Mol Res. 2014;13(1):799-805.
13. Aykut A, Onay H, Gunduz C, Ozkinay F, Cogulu O, Sagol S. Determination of fetal rhesus d status by maternal plasma DNA analysis. Balkan Journal of Medical Genetics. 2013;16(2):33-8.
14. Moezzi L, Keshavarz Z, Ranjbaran R, Aboualizadeh F, Behzad-Behbahani A, Abdullahi M, et al. Fetal RHD genotyping using real-time polymerase chain reaction analysis of cell-free fetal DNA in pregnancy of RhD negative women in South of Iran. International journal of fertility & sterility. 2016;10(1):62.
15. Addai-Mensah O, Afriyie EY, Sakyi SA, Obirikorang C, Annani-Akollor ME, Owiredu E-W, et al. Fetal Rhesus D genotyping and sex determination from maternal plasma of Rhesus D-negative antenatal population: the usefulness of conventional polymerase chain reaction in resource-limited settings. Obstetrics and Gynecology International. 2020;2020.
16. Bohmova J, Lubusky M, Holuskova I, Studnickova M, Kratochvilova R, Krejcirikova E, et al. Two reliable methodical approaches for non-invasive RHD genotyping of a fetus from maternal plasma. Diagnostics. 2020;10(8):564.
17. Haimila K, Sulin K, Kuosmanen M, Sareneva I, Korhonen A, Natunen S, et al. Targeted antenatal anti‐D prophylaxis program for RhD‐negative pregnant women–outcome of the first two years of a national program in Finland. Acta Obstetricia et Gynecologica Scandinavica. 2017;96(10):1228-33.
18. Hyland CA, Millard GM, O'Brien H, Schoeman EM, Lopez GH, McGowan EC, et al. Non-invasive fetal RHD genotyping for RhD negative women stratified into RHD gene deletion or variant groups: comparative accuracy using two blood collection tube types. Pathology. 2017;49(7):757-64.
19. Zhou L, Thorson JA, Nugent C, Davenport RD, Butch SH, Judd WJ. Noninvasive prenatal RHD genotyping by real-time polymerase chain reaction using plasma from D-negative pregnant women. American journal of obstetrics and gynecology. 2005;193(6):1966-71.
20. Bischoff FZ, Nguyen DD, Marquéz-Do D, Moise Jr KJ, Simpson JL, Elias S. Noninvasive determination of fetal RhD status using fetal DNA in maternal serum and PCR. Journal of the Society for Gynecologic Investigation. 1999;6(2):64-9.
21. Müller SP, Bartels I, Stein W, Emons G, Gutensohn K, Köhler M, et al. The determination of the fetal D status from maternal plasma for decision making on Rh prophylaxis is feasible. Transfusion. 2008;48(11):2292-301.
Published
2025-01-13
How to Cite
1.
Ahmad S, ABDULAZEEZ SALH S. FOETAL RHESUS D AND SEX GENOTYPING FROM THE PLASMAS OF RH-NEGATIVE PREGNANT WOMEN IN DUHOK. Duhok Medical Journal [Internet]. 13Jan.2025 [cited 13Jun.2026];18(2):1-0. Available from: https://dmj.uod.ac/index.php/dmj/article/view/224