A novel duplex SSP-PCR typing method for KIR gene profiling

• Published in: Tissue antigens 2009-07, Vol.74 (1), p.62-67
• Main Authors: Ashouri, E., Ghaderi, A., Reed, E. F., Rajalingam, R.
• Format: Article
• Language: English
• Subjects: Alleles; Genotype; Humans; Killer Cells, Natural - immunology; killer-cell immunoglobulin-like receptors; KIR gene profiling; KIR genotyping; natural killer-cell receptors; Polymerase Chain Reaction - methods; Receptors, KIR - genetics; Sensitivity and Specificity; Sequence Analysis, DNA - methods
• ISSN: 0001-2815; 1399-0039
• DOI: 10.1111/j.1399-0039.2009.01259.x

Killer‐cell immunoglobulin‐like receptors (KIR) control the function of natural killer cells. The number and type of KIR genes are substantially variable among individuals. Sequence‐specific primer–directed polymerase chain reaction (SSP‐PCR) based genotyping is the most commonly used method to assess the KIR gene content. However, it requires a minimum of 16 gene‐specific amplifications and often yields false‐negative results. Herein, we describe the development of a simple and efficient duplex SSP‐PCR assay to identify the presence and absence of 16 KIR genes. This system further distinguishes subsets of KIR2DS4 and KIR3DP1 alleles. The assay was subjected to a blind validation using a panel of 78 reference DNA standards from the UCLA KIR Exchange Program, which showed 100% specificity and accuracy. Compared with the conventional SSP typing methods, the present method is an accurate, simple, cost‐effective and labor‐saving KIR genotyping method for high volume testing.