Practical Considerations and Workflow in Utilizing KIR Genotyping in Transplantation Medicine

Yawata, Makoto; Yawata, Nobuyo

doi:10.1007/978-1-0716-2160-8_20

Cited by 2 publications

(2 citation statements)

References 51 publications

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“…PCR-SSP provide only 'presence/absence' status for a KIR gene, but it is still widely used in many research laboratories because it is well-established, most costeffective, and simple to perform. [35][36][37][38][39][40][41][42][43][44]58,59 The published PCR-SSP methods for KIR genotyping are simple and robust, but they have the drawback of using large amounts of input DNA (100 ng up to 1 μg per reaction) and being partially or totally based on the amplification of long DNA fragments that often span 0.5-2.0 kb. 10,11,35,38,54 These conditions require high-quality DNA; however, most clinical or anthropological studies may not provide sufficient amounts of high-quality DNA for a complete typing profile.…”

Section: Discussionmentioning

confidence: 99%

Development of cost‐effective and fast KIR genotyping by multiplex PCR‐SSP

Choi,

Baek,

Park

et al. 2023

HLA

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Killer‐cell immunoglobulin‐like receptors (KIR) control natural killer (NK) cell functions by recognizing HLA molecules and modulating the activity of NK cells. The KIR gene cluster contains polymorphic and highly homologous genes. Diversity of the KIR region is achieved through differences in gene content, allelic polymorphism, and gene copy number, which result in unrelated individuals having different KIR genotypes and individualized immune responses that are relevant to multiple aspects of human health and disease. Therefore, KIR genotyping is increasingly used in epidemiological studies. Here, we developed multiplex polymerase chain reaction with sequence‐specific primers (PCR‐SSP) to compensate for the shortcomings of the conventional PCR‐SSP method, which is most commonly used for KIR analysis. Multiplex PCR‐SSP method involves six multiplex reactions that detect 16 KIR genes and distinguish variant types of some KIR genes by adding two reactions. The assay was evaluated in a blind survey using a panel of 40 reference DNA standards from the UCLA KIR Exchange Program. The results are 100% concordant with the genotype determined using Luminex‐based reverse sequence‐specific oligonucleotide typing systems. Additionally, we investigated the currently known 16 KIR genes and their common variants in 120 unrelated Korean individuals. The results were consistent with the KIR genotype previously reported by Hwang et al. This multiplex PCR‐SSP is an efficient method for analyzing KIR genotypes in both small‐ and large‐scale studies with minimal labor, reagents, and DNA. Furthermore, by providing a better definition of KIR polymorphisms it can contribute to developments in immunogenetics.

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Section: Discussionmentioning

confidence: 99%

Development of cost‐effective and fast KIR genotyping by multiplex PCR‐SSP

Choi,

Baek,

Park

et al. 2023

HLA

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show abstract

“…76,77 KIR AND HLA GENOTYPING Methods to genotype HLA to allele level are well established and adopted by clinical laboratories worldwide, providing ample material for the multitude of candidate gene association studies. 34 Because structural complexity of the locus (Figure 2) has hindered similar attempts, gene presence/absence variation has been the method of choice 78 for most studies investigating KIR diversity in human health. Because gene-content variation is the major component of KIR-mediated NK cell functional diversity, these studies have been highly informative and generally reproducible but can be conflicting, particularly across ancestrydistinct cohorts.…”

Section: Kir Polymorphismmentioning

confidence: 99%