Patent ID: 9005894
Filing Date: 2015-04-14
Classification: C12Q,Y02A

Abstract:
1. A method of determining KIR genotypes by determining sequences of multiple exons in multiple KIR genes for one or more individuals in parallel, the method comprising: (a) for each individual, performing an amplification reaction for each KIR exon with a single forward primer and a single reverse primer, each primer comprising an adapter sequence, an individual identification sequence, and a KIR-hybridizing sequence hybridizing to and supporting amplification of the sequence from the same exon in more than one KIR gene, to amplify the exon sequences of more than one KIR genes that comprise polymorphic sites to obtain amplicons from more than one KIR gene; (b) pooling KIR amplicons from more than one individual obtained in step (a); (c) performing emulsion PCR; (d) determining the sequence of each KIR amplicon for each individual using pyrosequencing in parallel; and (e) assigning the KIR alleles to each individual by comparing the sequence of the KIR amplicons determined in step (d) to known KIR sequences to determine which KIR alleles are present in the individual thereby determining the KIR genotype.