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We have developed a new method for analysis of complex genetic systems by a combination of the Amplification Refractory Mutation System (ARMS) and Single-Strand Conformation Polymorphism (SSCP) analysis: ARMS-SSCP. Thus, a complex allelic series is subdivided into a number of groups by ARMS followed by the identification of specific alleles using SSCP analysis. We have shown that the HLA alleles at the DRB3 and DQB1 loci were distinguishable from each other using ARMS-SSCP. In 36 individuals typed for the DRB3 and 48 individuals typed for the DQB1 loci, ARMS-SSCP results were in complete agreement with those obtained using the established method of sequence-specific oligonucleotide (SSO) hybridisation. With silver staining, ARMS-SSCP is a rapid, non-radioactive and reliable method which also offers the possibility for detecting new HLA alleles. We have demonstrated that ARMS-SSCP can be performed using fluorescent PCR primers, a feature which gives the method potential for automation.

Original publication

DOI

10.1093/nar/20.5.1005

Type

Journal article

Journal

Nucleic acids research

Publication Date

03/1992

Volume

20

Pages

1005 - 1009

Addresses

Nuffield Department of Pathology and Bacteriology, John Radcliffe Hospital, Oxford, UK.

Keywords

Humans, DNA, Single-Stranded, Oligodeoxyribonucleotides, HLA-DQ Antigens, HLA-DR Antigens, Fluorescent Dyes, Silver Staining, Polymerase Chain Reaction, Genes, MHC Class II, Base Sequence, Nucleic Acid Conformation, Genotype, Mutation, Polymorphism, Genetic, Molecular Sequence Data, HLA-DQ beta-Chains, HLA-DRB3 Chains