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.
Gersuk
Vivian H
VH
Benaroya Research Institute at Virginia Mason, 1201 9th Ave., Seattle, WA 98101, USA. vgersuk@benaroyaresearch.org
Nepom
Gerald T
GT
D017931:DNA Primers; D018628:Gene Dosage; D015804:HLA-DR4 Antigen / Q000235:genetics*; D006801:Humans; D010641:Phenotype; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000191:economics / Q000379:methods*; D012680:Sensitivity and Specificity
D017931:DNA Primers; D015804:HLA-DR4 Antigen