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HLA‐DQA1 AND ‐DQB1 GENOTYPING BY PCR‐RFLP, HETERODUPLEX AND HOMODUPLEX ANALYSIS
Author(s) -
Teutsch S. M.,
Bennetts B. H.,
Castle M.,
Hibbins M.,
Heard R. N. S.,
Stewart G. J.
Publication year - 1996
Publication title -
international journal of immunogenetics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.41
H-Index - 47
eISSN - 1744-313X
pISSN - 1744-3121
DOI - 10.1111/j.1744-313x.1996.tb00272.x
Subject(s) - genotyping , heteroduplex , typing , restriction fragment length polymorphism , biology , polymerase chain reaction , genetics , allele , oligonucleotide , genotype , dna , gene
SUMMARY PCR‐RFLP typing methods for DQA1 and DQB1 in conjunction with the analysis of heteroduplex and homoduplex patterns have allowed a simple method for typing all of the major DQA1 and DQB1 alleles. This method has advantages over PCR amplification with sequence‐specific primers (PCR‐SSP), PCR hybridization with sequence‐specific oligonucleotide probes (PCR‐SSO) and other PCR‐RFLP strategies for typing DQ alleles. The analysis of heteroduplex and homoduplex patterns can be used in conjunction with other PCR typing systems such as PCR‐SSP as a confirmatory step with little additional work. In addition, a PCR‐RFLP strategy was designed for resolving the DQB 1*0602 and DQB 1 *0603 alleles, which involved the use of a primer containing a base mutation, creating a new restriction site which distinguished the two alleles. These techniques have enabled resolution of the major homozygous and heterozygous combinations of these DQA1 and DQB I alleles. The PCR‐RFLP technique does not require the large number of oligonucleotides that are necessary for both the PCR‐SSP and PCR‐SSO techniques and is thus both time and cost effective for infrequent or small numbers of samples.