This chapter will describe a method for KIR genotyping, which detects the presence or absence of each gene in a given individual, thus providing a KIR profile. The method involves gene-specific polymerase chain reaction (PCR) amplification using sequence-specific primers (PCR-SSP).

2017年6月1日 · The killer cell immunoglobulin-like receptor (KIR) region of human chromosome 19 contains up to 16 genes for natural killer (NK) cell receptors that...

2012年1月1日 · PCR with sequence-specific primers (SSP) is simplest and most widely used among techniques for studying KIR genotypes. Here, we present a protocol that details the critical steps of a method for KIR genotyping by PCR-SSP.

These methods overcome the tedious and expensive nature of conventional KIR genotyping and HLA-class I typing using sequence-specific priming (SSP) PCR, sequence-specific oligonucleotide (SSO) hybridisation or sequence-based typing (SBT).

KIR locus PCR primers: 10 µM of each oligonucleotide primer in water, store at −20°C. Table 1 describes the primer sets needed based on the presence or absence of specific KIR genes in the sample. Primers are listed in Table 2 (see Note 3)

We describe here a simple and reliable multiplex PCR-SSP (sequence-specific priming) method for relatively rapid and inexpensive genotyping of 15 KIR genes using standard agarose gel electrophoresis. Keywords: KIR genotyping, NK cell, KIR …

2012年10月25日 · Through the recognition of human leukocyte antigen (HLA)-I, KIR regulates the cytoactivity of the effector cell, begins the immunological process and regulates cytokine secretion. KIR gene clusters have extremely complex gene structures gradually formed from the single gene structure of KIR3DX1 (2).

In this study, we have successfully tested the feasibility of using whole genome amplification (WGA) to allow genotyping for killer cell immunoglobulin-like receptor (KIR) genes and human leucocyte antigen (HLA)-C KIR ligand dimorphism on HLA-C. WGA was achieved by multiple displacement amplification (MDA) using bacteriophage phi29 polymerase.

PCR with sequence-specific primers (SSP) is simplest and most widely used among techniques for studying KIR genotypes. Here, we present a protocol that details the critical steps of a method for KIR genotyping by PCR-SSP.

2006年10月11日 · In this study, a reverse transcriptase-polymerase chain reaction (RT-PCR) that uses new primer sets for the gene-specific detection of KIR transcripts is presented and validated. The key advantage of this RT-PCR method over previously published ones is that it was designed to detect transcripts of all confirmed allelic variants of the KIR genes ...