A real‐time PCR approach for rapid high resolution subtyping of HLA‐DRB1*04

We present a real‐time PCR approach for the identification and subtyping of HLA‐DR4 alleles. The technique, which uses sequence‐specific primers and probes in conjunction with real‐time PCR for the detection and differentiation of target alleles, is rapid, involves minimal hands‐on time, and is inexpensive compared to existing methods. Further, there is no post‐PCR handling, so the risk of contamination is avoided. We have validated the assay using 44 blinded and 56 unblinded samples, which were identified with 100% accuracy, sensitivity, and specificity. We demonstrate the applicability of this assay as an alternative approach to traditional HLA typing methods.