Blood group antigen profile predicted by molecular biology - use of real-time polymerase chain reaction to genotype important KEL, JK, RHD, and RHCE alleles

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Immunohematology

American National Red Cross

Subject: Medical Laboratory Technology

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ISSN: 0894-203X
eISSN: 1930-3955

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VOLUME 18 , ISSUE 3 (September 2002) > List of articles

Blood group antigen profile predicted by molecular biology - use of real-time polymerase chain reaction to genotype important KEL, JK, RHD, and RHCE alleles

Fernando Manuel Ferreira Araújo / Christiana Pereira / Fátima Monteiro / Isabel Henriques / Elsa Meireles / Pedro Lacerda / Ana Aleixo / Regina Celeste / Luis M. Cunha-Ribeiro / Maria J. Rodrigues

Keywords : RHD, RHCE, KEL, JK, real-time PCR, LightCycler

Citation Information : Immunohematology. Volume 18, Issue 3, Pages 59-64, DOI: https://doi.org/10.21307/immunohematology-2019-511

License : (Transfer of Copyright)

Published Online: 14-October-2020

ARTICLE

ABSTRACT

The most clinically important blood group systems in transfusion medicine, excluding the ABO system, are the RH, Kell, and Kidd systems. Alloantibodies to antigens of these systems may be produced following blood transfusion or during pregnancy and can result in serious hemolytic transfusion reactions and hemolytic disease of the newborn.We developed rapid and robust techniques for RHD, RHCE, KEL, and JK genotyping with the use of a real-time polymerase chain reaction instrument. Two fluorescence-based methods for the detection of amplification products were used: for KEL1/KEL2, JK1/JK2, and RHE/RHe (exon 5) we used the hybridization probes protocol; for RHC/RHc the analysis was done in sequences of exon 1 for RHC and exon 2 for RHc; and for RHD, analysis was done in sequences of intron 4, exon 7, and exon 4 pseudogene using the SYBR Green I protocol. The genotyping tests were validated with samples from 85 Caucasian Portuguese and 15 Black European blood donors. Complete phenotype-genotype correlations were obtained. The potential use of the presented methods can be predicted in clinical transfusion medicine,allowing appropriate monitoring, early intervention, and improved care. When blood group genotyping techniques are necessary, this methodology is highly competitive for a routine laboratory.

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