117-P : KIR TYPING WITH LINKSEQ™, A REAL-TIME PCR DETECTION SYSTEM

Aim Killer cell immunoglobulin-like receptors (KIR) are a family of cell surface receptors found on human Natural Killer (NK) cells. Certain KIR types stimulate the activity of NK cells while others have been shown to interact with specific Human Leukocyte Antigen (HLA) Class I molecules to confer an inhibitory effect. This regulation of NK cell effector function has major implications for the role of HLA/KIR interactions in autoimmunity, viral progression, tumor immunity, and hematopoietic stem cell transplantation. Current KIR genotyping methods generally require a labor intensive process of running agarose gels or setting up SSOP reactions. Our goal in this study was to apply the LinkSeq method to reduce the hands on laboratory time of current methods and provide an automated method of analysis, thereby increasing laboratory productivity. Methods We developed 23 assays that together provide tests for a) all of the major KIR gene groups, b) the full length and deleted forms of KIR2DS4, KIR3DP1, and c) the specific alleles 3DL1004 and the aa245 variants of the KIR2DL1 group. In addition, software analysis features were included in SureTyper™ to automate genotyping and simplify reporting. Results This new KIR typing system reduces total test time by 50% and hands-on time by up to 90% compared to current methods, with an added benefit of reducing DNA input requirements by 1/10th compared to SSP methods. With less than 10 minutes of set-up and no further operator intervention, LinkSeq uses state of the art real-time PCR detection combined with melt curve analysis to provide specific amplicon identification. Further, since amplified products are never handled, the risk of laboratory contamination is avoided. Conclusions Given the ease of use, rapid turnaround times, and increased laboratory productivity, KIR typing with LinkSeq offers an attractive improvement over current SSP and SSOP typing methods. Russnak: Linkage Biosciences: Consultant. King: Linkage Biosciences: Employee; Stockholder. Ly: Linkage Biosciences: Employee; Stockholder. Quinto: Linkage Biosciences: Employee. Antovich: Linkage Biosciences: Employee; Stockholder.