Validation of a Polymerase Chain Reaction technique for Kidd blood group genotyping

Abstract

The Kidd blood group antigens, Jkª and Jkᵇ , are two of the main surface markers which are found on the membrane of red blood cells. The determination of whether a donor or a recipient has the Jkª and/or the Jkᵇ antigens is crucially important to have a successful transfusion without the development of adverse incompatibility-related reactions. In Malta, routine serological-based tests are applied with the purpose of differentiating between homozygous and heterozygous states for the Jk antigens respectively. Although these tests are highly specific and sensitive, there are particular clinical scenarios where haemagglutination assays are not suitable for determining the individual’s Kidd blood group status. Additionally, the alternative genotyping procedure has never been applied in Malta within the context of blood grouping. The current study was therefore carried out to determine whether a molecular-based technique such as Polymerase Chain Reaction – Restriction Fragment Length Polymorphism analysis (PCR-RFLP) is a suitable alternative procedure for distinguishing amongst the three different Kidd phenotypes. After extracting deoxyribonucleic acid (DNA) from 50 blood samples obtained from serologically-tested healthy blood donors who expressed at least one of the Kidd antigens, PCR-RFLP analyses were carried out. The results of the latter were then compared with those previously obtained with haemagglutination and a complete match was observed between the two. Therefore, this PCR-RFLP method was confirmed as a suitable alternative laboratory technique that can be used to determine efficiently the Kidd blood group of both donors and recipients, in an accurate manner without subjectivity as encountered in the case of haemagglutination. This research further facilitates the introduction of molecular-based techniques in molecular blood transfusion.