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High-throughput genotyping of KIR2DL2/L3, KIR3DL1/S1, and their HLA class I ligands using real-time PCR
Authors:R. N. Koehler,A. M. Walsh,N. Moqueet,J. R. Currier,M. A. Eller,L. A. Eller,F. Wabwire-Mangen,N. L. Michael,M. L. Robb,F. E. McCutchan,&   G. H. Kijak
Affiliation:Division of Retrovirology, US Military HIV Research Program/Henry M. Jackson Foundation, Rockville, MD, USA;
Makerere University Walter Reed Project, Henry M. Jackson Foundation, Kampala, Uganda;
Makerere University, Kampala, Uganda;
Division of Retrovirology, US Military HIV Research Program/Walter Reed Army Institute of Research, Silver Spring, MD, USA
Abstract:Killer immunoglobulin-like receptors (KIRs) expressed on natural killer cells are critical components of innate immunity. Interactions between KIRs and their human leukocyte antigen (HLA) ligands have been shown to influence autoimmune and infectious disease course in defined populations. However, the low throughput and high cost of current methods impede confirmation of the universality of these findings. To support large epidemiology surveys, we developed a high-throughput real-time polymerase chain reaction-based assay to identify carriers of KIR3DL1, KIR3DS1, KIR2DL2, and KIR2DL3 and their HLA ligands. The platform performed with 100% sensitivity and specificity in detection of carrier and non-carrier on reference samples. The application of this platform will further clarify the nature and impact of the KIR–HLA epistatic interaction on disease course in large global population-based studies.
Keywords:human leukocyte antigen    innate immunity    killer immunoglobulin-like receptor    real-time polymerase chain reaction    TaqMan
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