[en] OBJECTIVES: In some European countries, non-invasive fetal RHD genotyping is the first step of anti-D allo-immunized pregnant women management but presence of RHD variant alleles may interfere with the results accuracy. We developed an algorithm allowing solving discordant results (due to the presence of RHD variant) in fetal RHD genotyping assay.
METHOD: This study gathered the results of fetal RHD genotyping performed between 2006 and 2020 in the Medicine Laboratory of CHR Liège. Exons 4, 5 and 10 of the fetal RHD were profiled in maternal plasma using real time polymerase chain reaction (PCR). When the results were discrepant, maternal RHD variant was further explored by sequence-specific primer PCR on maternal buffy coat.
RESULTS: A total of 11,630 pregnant women (mainly of both Caucasian and African origins) were tested during the study period and RHD variant alleles were detected in 247 women. The most frequent variant was RHD*08N.01 found in 66 women mainly of Black African origin. We identified 45 women with weak RHD variant type 1, 2 or 3.
CONCLUSION: Women with weak RHD variant type 1, 2 or 3 can safely be considered as RhD positive in terms of RhIg prophylaxis and/or transfusion of blood components. Therefore, identification of RHD allele variants in women with discordant fetal RHD genotyping results contributes to save RhIg prophylaxis and RhD negative blood components.
Disciplines :
Laboratory medicine & medical technology Reproductive medicine (gynecology, andrology, obstetrics)
Author, co-author :
Dufour, Patrice ; Centre Hospitalier Universitaire de Liège - CHU > > Pool assistant - biologie clinique ; Department of Laboratory Medicine, Immunohaematology and Transfusion Medicine Division, Centre Hospitalier Régional de la Citadelle, Liège, Belgium
Gerard, Christiane; Blood Transfusion Service, University of Liège (ULiège), CHU, Liège, Belgium
Chantraine, Frédéric ; Centre Hospitalier Universitaire de Liège - CHU > > Service de gynécologie-obstétrique (CHR)
Minon, Jean-Marc; Department of Laboratory Medicine, Immunohaematology and Transfusion Medicine Division, Centre Hospitalier Régional de la Citadelle, Liège, Belgium
Language :
English
Title :
Investigation of discrepancies obtained during 15 years of non-invasive fetal RHD genotyping in apparent serologic RhD-negative pregnant women.
Toly-Ndour C, Huguet-Jacquot S, Mailloux A, et al. Rh disease prevention: the European Perspective. ISBT Sci Ser. 2021;16(1):106-118. https://doi.org/10.1111/voxs.12617
van der Schoot CE, de Haas M, Clausen FB. Genotyping to prevent Rh disease: has the time come? Curr Opin Hematol. 2017;24(6):544-550. https://doi.org/10.1097/moh.0000000000000379
Flegel WA, Wagner FF. Molecular biology of partial D and weak D: implications for blood bank practice. Clin Lab. 2002;48(1-2):53-59.
Judd WJ. Scientific Section Coordinating Committee of the AABB. Practice guidelines for prenatal and perinatal immunohematology, revisited. Transfusion. 2001;41(11):1445-1452. https://doi.org/10.1046/j.1537-2995.2001.41111445.x
Singleton BK, Green CA, Avent ND, et al. The presence of an RHD pseudogene containing a 37 base pair duplication and a nonsense mutation in africans with the Rh D-negative blood group phenotype. Blood. 2000;95(1):12-18. https://doi.org/10.1182/blood.v95.1.12
Manfroi S, Calisesi C, Fagiani P, et al. Prenatal non-invasive foetal RHD genotyping: diagnostic accuracy of a test as a guide for appropriate administration of antenatal anti-D immunoprophylaxis. Blood Transfus. 2018;16(6):514-524.
Minon JM, Gerard C, Senterre JM, Schaaps JP, Foidart JM. Routine fetal RHD genotyping with maternal plasma: a four-year experience in Belgium. Transfusion. 2008;48(2):373-381.
ISBT. Red Cell Immunogenetics and Blood Group Terminology. ISBT Working Party [Internet]. Retrieved March 23, 2022, from https://www.isbtweb.org/isbt-working-parties/rcibgt.html
Wagner FF, Kasulke D, Kerowgan M, Flegel WA. Frequencies of the blood groups ABO, Rhesus, D category VI, Kell, and of clinically relevant high-frequency antigens in south-western Germany. Infusionsther Transfusionsmed. 1995;22(5):285-290. https://doi.org/10.1159/000223144
Ansart-Pirenne H, Asso-Bonnet M, Le Pennec PY, Roussel M, Patereau C, Noizat-Pirenne F. RhD variants in Caucasians: consequences for checking clinically relevant alleles. Transfusion. 2004;44(9):1282-1286. https://doi.org/10.1111/j.1537-2995.2004.04063.x
Sandler SG, Chen LN, Flegel WA. Serological weak D phenotypes: a review and guidance for interpreting the RhD blood type using the RHD genotype. Br J Haematol. 2017;179(1):10-19. https://doi.org/10.1111/bjh.14757
Flegel WA. Molecular genetics and clinical applications for RH. Transfus Apher Sci. 2011;44(1):81-91. https://doi.org/10.1016/j.transci.2010.12.013
Virk M, Sandler SG. Rh immunoprophylaxis for women with a serologic weak D phenotype. Lab Med. 2015;46(3):190-194. https://doi.org/10.1309/lmunup4fjtux2gcd
Flegel WA, Peyrard T, Chiaroni J, Tournamille C, Jamet D, Pirenne F. A proposal for a rational transfusion strategy in patients of European and North African descent with weak D type 4.0 and 4.1 phenotypes. Blood Transfus. 2019;17(2):89-90.
Clausen FB, Steffensen R, Christiansen M, et al. Routine noninvasive prenatal screening for fetal RHD in plasma of RhD-negative pregnant women-2 years of screening experience from Denmark. Prenat Diagn. 2014;34(10):1000-1005. https://doi.org/10.1002/pd.4419
Haimila K, Sulin K, Kuosmanen M, 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 Obstet Gynecol Scand. 2017;96(10):1228-1233. https://doi.org/10.1111/aogs.13191
Stegmann TC, Veldhuisen B, Bijman R, et al. Frequency and characterization of known and novel RHD variant alleles in 37 782 Dutch D-negative pregnant women. Br J Haematol. 2016;173(3):469-479. https://doi.org/10.1111/bjh.13960
Clausen FB, Hellberg Å, Bein G, et al. Recommendation for validation and quality assurance of non-invasive prenatal testing for foetal blood groups and implications for IVD risk classification according to EU regulations. Vox Sang. 2022;117(2):157-165. https://doi.org/10.1111/vox.13172
Chitty LS, Finning K, Wade A, et al. Diagnostic accuracy of routine antenatal determination of fetal RHD status across gestation: population based cohort study. BMJ [Internet]. 2014;349:g5243. https://doi.org/10.1136/bmj.g5243
Moise KJ, Gandhi M, Boring NH, et al. Circulating cell-free DNA to determine the fetal RHD status in all three trimesters of pregnancy. Obstet Gynecol. 2016;128(6):1340-1346. https://doi.org/10.1097/aog.0000000000001741
Soothill PW, Finning K, Latham T, Wreford-Bush T, Ford J, Daniels G. Use of cffDNA to avoid administration of anti-D to pregnant women when the fetus is RhD-negative: implementation in the NHS. BJOG. 2015;122(12):1682-1686. https://doi.org/10.1111/1471-0528.13055
Vivanti A, Benachi A, Huchet FX, Ville Y, Cohen H, Costa JM. Diagnostic accuracy of fetal rhesus D genotyping using cell-free fetal DNA during the first trimester of pregnancy. Am J Obstet Gynecol. 2016;215(5):606.e1-606.e5. https://doi.org/10.1016/j.ajog.2016.06.054
Finning K, Martin P, Summers J, Massey E, Poole G, Daniels G. Effect of high throughput RHD typing of fetal DNA in maternal plasma on use of anti-RhD immunoglobulin in RhD negative pregnant women: prospective feasibility study. BMJ. 2008;336(7648):816-818. https://doi.org/10.1136/bmj.39518.463206.25
Wikman AT, Tiblad E, Karlsson A, Olsson ML, Westgren M, Reilly M. Noninvasive single-exon fetal RHD determination in a routine screening program in early pregnancy. Obstetrics Gynecol. 2012;120(2 Part 1):227-234. https://doi.org/10.1097/aog.0b013e31825d33d9
Ziza KC, Liao AW, Dezan M, et al. Determination of fetal RHD genotype including the RHD pseudogene in maternal plasma. J Clin Lab Anal. 2017;31(3):e22052. https://doi.org/10.1002/jcla.22052
Rouillac-Le Sciellour C, Puillandre P, Gillot R, et al. Large-scale pre-diagnosis study of fetal RHD genotyping by PCR on plasma DNA from RhD-negative pregnant women. Mol Diagn. 2004;8(1):23-31. https://doi.org/10.1007/bf03260044
Müller SP, Bartels I, Stein W, et al. The determination of the fetal D status from maternal plasma for decision making on Rh prophylaxis is feasible. Transfusion. 2008;48(11):2292-2301. https://doi.org/10.1111/j.1537-2995.2008.01843.x
Akolekar R, Finning K, Kuppusamy R, Daniels G, Nicolaides KH. Fetal RHD genotyping in maternal plasma at 11-13 weeks of gestation. Fetal Diagn Ther. 2011;29(4):301-306. https://doi.org/10.1159/000322959
Boggione CT, Luján Brajovich ME, Mattaloni SM, et al. Genotyping approach for non-invasive foetal RHD detection in an admixed population. Blood Transfus. 2017;15(1):66-73.
Parchure D, Madkaikar M, Kulkarni S. Algorithm development and diagnostic accuracy testing for non-invasive foetal RHD genotyping: an Indian experience. Blood Transfus. 2022;20(3):235-244.
Hyland CA, Gardener GJ, O’Brien H, et al. Strategy for managing maternal variant RHD alleles in Rhesus D negative obstetric populations during fetal RHD genotyping. Prenat Diagn. 2014;34(1):56-62. https://doi.org/10.1002/pd.4253
Power JP, Lawlor E, Davidson F, et al. Hepatitis C viraemia in recipients of Irish intravenous anti-D immunoglobulin. Lancet. 1994;344(8930):1166-1167. https://doi.org/10.1016/s0140-6736(94)90679-3
