A comparison between using Peripheral Whole Blood and Buccal Swab For Red Cell Genotyping amongst Multiply-Transfused Thalassaemia patients

• Main Author: Nadila Haryani Osman
• Format: Thesis
• Language: en_US
• Published: Universiti Sains Islam malaysia 2016
• Subjects: Thalassemia
• Online Access: http://ddms.usim.edu.my:80/jspui/handle/123456789/12502

Blood has always been the main sample in determining the different blood group types. However, in multiply-transfused patients, accurate red cell antigen typing by serology is a constant problem due to the exposure to donor‟s blood in patient‟s circulation making results unreliable. To overcome these problems, another source and technique for blood typing is needed. The main aim of this study was to investigate the genotype of red cells for RH (C, c, E, e), KEL (Kell, Celano), Kidd (JKA, JKB) and Duffy (FYA, FYB) using buccal swab and peripheral whole blood amongst multiply-transfused thalassaemia patients. Sixty-three of multiply-transfused thalassaemia patients from the Thalassaemia Clinic of Ampang Hospital and Universiti Kebangsaan Malaysia Medical Centre participated in this study. Paired samples consisting of peripheral whole blood and buccal swab samples were collected prior to the scheduled blood transfusion and on day 7 after the transfusion. Blood samples were subjected to serological phenotyping by tube method and DNA genotyping while buccal swab was subjected to DNA genotyping only. Blood genotyping was performed using TaqMan® Single Nucleotide Polymorphism Real Time Polymerase Chain Reaction (SNP RT-PCR) assays for RHEe, RHCc, Kidd, KEL and Duffy blood group systems. Complete data was available in 33 patients only. Discrepancies were found between the phenotype and genotype results for all blood groups tested in both pre- and post-transfusion samples. However, a full concordance of genotyping result between pre- and post-transfusion samples was observed. When comparing the genotyping results between different sampling methods, blood and buccal swab samples showed concordant results. Accurate red blood cell antigen profiling is important for patients requiring multiple transfusions. The SNP RT-PCR platform is a reliable alternative to the conventional method. Buccal samples offer a simple and inexpensive alternative collection method that may be used for accurate blood group genotyping when blood samples are unavailable.