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Candidate Gene Testing in Clinical Cohort Studies with Multiplexed Genotyping and Mass Spectrometry
Published on: June 21, 2018
An Easy Multiplex PCR-SSP Assay for the Genotyping of KEL1 and KEL2 in Multi-transfused Patients
Parinaz Zarghamian1, Maryam Pourshadlou1, Kamran Mousavi Hosseini1
1Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Hemmat Expressway, IBTO Building, Tehran, 1449613111 Iran.
Abstract:
Kell blood group system consists of 34 antigens. KEL1 and KEL2 are the most clinically important antigens of this system, causing hemolytic disease of the fetus and newborn (HDFN) and transfusion reaction. A total of 200 samples from blood donors were tested serologically for the presence of KEL1 and KEL2 antigens on erythrocytes. Genomic DNA was analyzed by PCR-SSP method to determine the Kell genotype. A multiplex PCR-SSP assay was designed and tested to genotype KEL1/KEL2 alleles in a single reaction. PCR genotyping revealed samples as; KEL2/KEL2 (93.5%) and KEL1/KEL2 (6.5%), while no sample determined as KEL1/KEL1. A 100% concordance observed between PCR and serological results. Multiplex PCR accurately diagnosed Kell genotype. Kell blood group genotyping by PCR-SSP can be used as an alternative method, especially in multi-transfused patients where serological findings are ambiguous.

