Design and validation of a multiplex specific primer-directed polymerase chain reaction assay for killer-cell immunoglobulin-like receptor genetic profiling

A. T. Abalos; R. Eggers; M. Hogan; C. M. Nielson; A. R. Giuliano; R. B. Harris; P. A. Thompson

doi:10.1111/j.1399-0039.2010.01588.x

Design and validation of a multiplex specific primer-directed polymerase chain reaction assay for killer-cell immunoglobulin-like receptor genetic profiling

A. T. Abalos, R. Eggers, M. Hogan, C. M. Nielson, A. R. Giuliano, R. B. Harris, P. A. Thompson

Research output: Contribution to journal › Article › peer-review

8 Scopus citations

Abstract

Current methodologies for the analysis of the killer-cell immunoglobulin-like receptor (KIR) locus utilize specific primer-directed polymerase chain reaction (SSP-PCR), which require a wide range of DNA input, multiple reaction conditions, and up to 16 individual reactions. We have developed and validated a multiplex SSP-PCR method for the genetic analysis of the KIR locus. Design and optimization of four multiplex groups targeting 14 genes and their alleles on the KIR locus has been completed. Each multiplex group contains PCR products that differ in size by a minimum of 15 bp to allow sufficient fragment length resolution for size discrimination by gel electrophoresis. This assay allows for efficient genotyping of the KIR locus while requiring a minimum amount of DNA input, utilizing the simplicity of SSP-PCR.

Original language	English (US)
Pages (from-to)	143-148
Number of pages	6
Journal	Tissue Antigens
Volume	77
Issue number	2
DOIs	https://doi.org/10.1111/j.1399-0039.2010.01588.x
State	Published - Feb 2011
Externally published	Yes

Keywords

Killer-cell immunoglobulin-like receptor genotyping
Killer-cell immunoglobulin-like receptors
Multiplex polymerase chain reaction
Specific primer-directed polymerase chain reaction

ASJC Scopus subject areas

Immunology and Allergy
Immunology
Biochemistry
Genetics

Access to Document

10.1111/j.1399-0039.2010.01588.x

Cite this

@article{e8c6fa70fcd84710b48aa1144f16a9ee,

title = "Design and validation of a multiplex specific primer-directed polymerase chain reaction assay for killer-cell immunoglobulin-like receptor genetic profiling",

abstract = "Current methodologies for the analysis of the killer-cell immunoglobulin-like receptor (KIR) locus utilize specific primer-directed polymerase chain reaction (SSP-PCR), which require a wide range of DNA input, multiple reaction conditions, and up to 16 individual reactions. We have developed and validated a multiplex SSP-PCR method for the genetic analysis of the KIR locus. Design and optimization of four multiplex groups targeting 14 genes and their alleles on the KIR locus has been completed. Each multiplex group contains PCR products that differ in size by a minimum of 15 bp to allow sufficient fragment length resolution for size discrimination by gel electrophoresis. This assay allows for efficient genotyping of the KIR locus while requiring a minimum amount of DNA input, utilizing the simplicity of SSP-PCR.",

keywords = "Killer-cell immunoglobulin-like receptor genotyping, Killer-cell immunoglobulin-like receptors, Multiplex polymerase chain reaction, Specific primer-directed polymerase chain reaction",

author = "Abalos, {A. T.} and R. Eggers and M. Hogan and Nielson, {C. M.} and Giuliano, {A. R.} and Harris, {R. B.} and Thompson, {P. A.}",

year = "2011",

month = feb,

doi = "10.1111/j.1399-0039.2010.01588.x",

language = "English (US)",

volume = "77",

pages = "143--148",

journal = "Tissue Antigens",

issn = "0001-2815",

publisher = "Wiley-Blackwell",

number = "2",

}

TY - JOUR

T1 - Design and validation of a multiplex specific primer-directed polymerase chain reaction assay for killer-cell immunoglobulin-like receptor genetic profiling

AU - Abalos, A. T.

AU - Eggers, R.

AU - Hogan, M.

AU - Nielson, C. M.

AU - Giuliano, A. R.

AU - Harris, R. B.

AU - Thompson, P. A.

PY - 2011/2

Y1 - 2011/2

N2 - Current methodologies for the analysis of the killer-cell immunoglobulin-like receptor (KIR) locus utilize specific primer-directed polymerase chain reaction (SSP-PCR), which require a wide range of DNA input, multiple reaction conditions, and up to 16 individual reactions. We have developed and validated a multiplex SSP-PCR method for the genetic analysis of the KIR locus. Design and optimization of four multiplex groups targeting 14 genes and their alleles on the KIR locus has been completed. Each multiplex group contains PCR products that differ in size by a minimum of 15 bp to allow sufficient fragment length resolution for size discrimination by gel electrophoresis. This assay allows for efficient genotyping of the KIR locus while requiring a minimum amount of DNA input, utilizing the simplicity of SSP-PCR.

AB - Current methodologies for the analysis of the killer-cell immunoglobulin-like receptor (KIR) locus utilize specific primer-directed polymerase chain reaction (SSP-PCR), which require a wide range of DNA input, multiple reaction conditions, and up to 16 individual reactions. We have developed and validated a multiplex SSP-PCR method for the genetic analysis of the KIR locus. Design and optimization of four multiplex groups targeting 14 genes and their alleles on the KIR locus has been completed. Each multiplex group contains PCR products that differ in size by a minimum of 15 bp to allow sufficient fragment length resolution for size discrimination by gel electrophoresis. This assay allows for efficient genotyping of the KIR locus while requiring a minimum amount of DNA input, utilizing the simplicity of SSP-PCR.

KW - Killer-cell immunoglobulin-like receptor genotyping

KW - Killer-cell immunoglobulin-like receptors

KW - Multiplex polymerase chain reaction

KW - Specific primer-directed polymerase chain reaction

UR - http://www.scopus.com/inward/record.url?scp=78650932541&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=78650932541&partnerID=8YFLogxK

U2 - 10.1111/j.1399-0039.2010.01588.x

DO - 10.1111/j.1399-0039.2010.01588.x

M3 - Article

C2 - 21214526

AN - SCOPUS:78650932541

SN - 0001-2815

VL - 77

SP - 143

EP - 148

JO - Tissue Antigens

JF - Tissue Antigens

IS - 2

ER -

Design and validation of a multiplex specific primer-directed polymerase chain reaction assay for killer-cell immunoglobulin-like receptor genetic profiling

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this