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HLA‐ DQB1 AND DQA1 MATCHING BY AMBIENT TEMPERATURE PCR‐SSCP
Author(s) -
Clay T.M.,
Culpan D.,
Pursall M.C.,
Bradley B.A.,
Bidwell J.L.
Publication year - 1995
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.1995.tb00283.x
Subject(s) - single strand conformation polymorphism , polymerase chain reaction , microbiology and biotechnology , biology , restriction fragment length polymorphism , polymorphism (computer science) , genetics , allele , gene
SUMMARY We have developed a polymerase chain reaction‐single strand conformation polymorphism (PCR‐SSCP) protocol for rapid matching of DQA1 and DQB1 alleles. Electrophoresis can be performed at ambient temperature within the range 18‐28°C without continuous gel cooling. The method has been tested on 27 patient‐potential bone marrow donor pairs for DQB1 and 31 pairs for DQA1. Bone marrow pairs were chosen to represent a broad range of common alleles based upon previous restriction fragment length polymorphism (RFLP) analysis type assignments. Samples were re‐typed by PCR with sequence‐specific primers (PCR‐SSP) and the results compared to matching by PCR‐SSCP analysis. There was a 100% correlation between PCR‐SSP and PCR‐SSCP analysis for DQB1 , and a 97% correlation for DQAl matching.