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

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 asse...

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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
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cited_by cdi_FETCH-LOGICAL-c5029-2e206a4d40ac51054790f9868469b57b2b568992dfdb9a353182ee6eb33ed40b3
cites cdi_FETCH-LOGICAL-c5029-2e206a4d40ac51054790f9868469b57b2b568992dfdb9a353182ee6eb33ed40b3
container_end_page 67
container_issue 1
container_start_page 62
container_title Tissue antigens
container_volume 74
creator Ashouri, E.
Ghaderi, A.
Reed, E. F.
Rajalingam, R.
description 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.
doi_str_mv 10.1111/j.1399-0039.2009.01259.x
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source Wiley-Blackwell Read & Publish Collection
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
title A novel duplex SSP-PCR typing method for KIR gene profiling
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