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Abstract
Genotyping has become an indispensable technique widely used in transfusion medicine. However, further simplification of the process is essential. Among the simplest methods, TaqMan-PCR is prominent. Nevertheless, challenges related to time, cost and the risk of sample mix-up during the DNA extraction process persist. To address these challenges, we have developed a novel blood processing method. Whole blood collected from healthy adult blood donors was heated at 95°C for 5 min and then used as a TaqMan-PCR template. The human platelet antigen (HPA)-1, -2, -3, -4, -5 and -15 genotypes of 60 individuals were evaluated. The results were fully consistent with those obtained through routine testing using PCR with reverse-sequence-specific oligonucleotide (rSSO) hybridization, confirming the reliability of our approach. The developed 'one-tube method' significantly reduces the risk of sample mix-up while offering considerable time and cost savings. Furthermore, this method has broader applicability to other genes, as the polymorphic regions tested in this study exhibited variability in guanine-cytosine content and amplicon lengths.
Published Version
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