Allelic typing of the HLA-DR4 group by polymerase chain reaction: single-strand conformation polymorphism analysis

Neil Thomas Young; C Darke

doi:10.1016/0198-8859(93)90144-P

Allelic typing of the HLA-DR4 group by polymerase chain reaction: single-strand conformation polymorphism analysis

Neil Thomas Young, C Darke

Applied Medicine

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

14 Citations (Scopus)

Abstract

The technique of SSCP was applied to the molecular subtyping of the HLA-DR4 group of alleles. SSCPs were defined in 16 cell line DNAs, representing the alleles DRB1*0401-*0411 (except DRB1*0409), using group-specific PCR amplification, nondenaturing polyacrylamide gel electrophoresis, and silver staining. Three sets of gel electrophoresis conditions were necessary to differentiate the 10 alleles tested. The SSCPs defined in the cell line DNAs were validated by analysis of 145 unrelated HLA-DR4-positive individuals previously genotyped for DRB1*04 alleles by sequence-specific oligotyping. This study demonstrates the suitability of the SSCP technique for defining DRB1*04 alleles, particularly when a small number of samples require typing.

Original language	English
Pages (from-to)	69-74
Number of pages	6
Journal	Human Immunology
Volume	37
Issue number	2
DOIs	https://doi.org/10.1016/0198-8859(93)90144-P
Publication status	Published - 1 Jun 1993

Keywords

Alleles
Base Sequence
DNA, Single-Stranded
Electrophoresis, Polyacrylamide Gel
Genotype
HLA-DR4 Antigen
Humans
Molecular Sequence Data
Oligonucleotide Probes
Polymerase Chain Reaction
Polymorphism, Genetic
Reproducibility of Results
Sensitivity and Specificity

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10.1016/0198-8859(93)90144-P

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title = "Allelic typing of the HLA-DR4 group by polymerase chain reaction: single-strand conformation polymorphism analysis ",

abstract = "The technique of SSCP was applied to the molecular subtyping of the HLA-DR4 group of alleles. SSCPs were defined in 16 cell line DNAs, representing the alleles DRB1*0401-*0411 (except DRB1*0409), using group-specific PCR amplification, nondenaturing polyacrylamide gel electrophoresis, and silver staining. Three sets of gel electrophoresis conditions were necessary to differentiate the 10 alleles tested. The SSCPs defined in the cell line DNAs were validated by analysis of 145 unrelated HLA-DR4-positive individuals previously genotyped for DRB1*04 alleles by sequence-specific oligotyping. This study demonstrates the suitability of the SSCP technique for defining DRB1*04 alleles, particularly when a small number of samples require typing.",

keywords = "Alleles, Base Sequence, DNA, Single-Stranded, Electrophoresis, Polyacrylamide Gel, Genotype, HLA-DR4 Antigen, Humans, Molecular Sequence Data, Oligonucleotide Probes, Polymerase Chain Reaction, Polymorphism, Genetic, Reproducibility of Results, Sensitivity and Specificity",

author = "Young, {Neil Thomas} and C Darke",

year = "1993",

month = jun,

day = "1",

doi = "10.1016/0198-8859(93)90144-P",

language = "English",

volume = "37",

pages = "69--74",

journal = "Human Immunology",

issn = "0198-8859",

publisher = "Elsevier Inc.",

number = "2",

}

TY - JOUR

T1 - Allelic typing of the HLA-DR4 group by polymerase chain reaction

T2 - single-strand conformation polymorphism analysis

AU - Young, Neil Thomas

AU - Darke, C

PY - 1993/6/1

Y1 - 1993/6/1

N2 - The technique of SSCP was applied to the molecular subtyping of the HLA-DR4 group of alleles. SSCPs were defined in 16 cell line DNAs, representing the alleles DRB1*0401-*0411 (except DRB1*0409), using group-specific PCR amplification, nondenaturing polyacrylamide gel electrophoresis, and silver staining. Three sets of gel electrophoresis conditions were necessary to differentiate the 10 alleles tested. The SSCPs defined in the cell line DNAs were validated by analysis of 145 unrelated HLA-DR4-positive individuals previously genotyped for DRB1*04 alleles by sequence-specific oligotyping. This study demonstrates the suitability of the SSCP technique for defining DRB1*04 alleles, particularly when a small number of samples require typing.

AB - The technique of SSCP was applied to the molecular subtyping of the HLA-DR4 group of alleles. SSCPs were defined in 16 cell line DNAs, representing the alleles DRB1*0401-*0411 (except DRB1*0409), using group-specific PCR amplification, nondenaturing polyacrylamide gel electrophoresis, and silver staining. Three sets of gel electrophoresis conditions were necessary to differentiate the 10 alleles tested. The SSCPs defined in the cell line DNAs were validated by analysis of 145 unrelated HLA-DR4-positive individuals previously genotyped for DRB1*04 alleles by sequence-specific oligotyping. This study demonstrates the suitability of the SSCP technique for defining DRB1*04 alleles, particularly when a small number of samples require typing.

KW - Alleles

KW - Base Sequence

KW - DNA, Single-Stranded

KW - Electrophoresis, Polyacrylamide Gel

KW - Genotype

KW - HLA-DR4 Antigen

KW - Humans

KW - Molecular Sequence Data

KW - Oligonucleotide Probes

KW - Polymerase Chain Reaction

KW - Polymorphism, Genetic

KW - Reproducibility of Results

KW - Sensitivity and Specificity

U2 - 10.1016/0198-8859(93)90144-P

DO - 10.1016/0198-8859(93)90144-P

M3 - Article

C2 - 8226138

SN - 0198-8859

VL - 37

SP - 69

EP - 74

JO - Human Immunology

JF - Human Immunology

IS - 2

ER -

Allelic typing of the HLA-DR4 group by polymerase chain reaction: single-strand conformation polymorphism analysis

Abstract

Keywords

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