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COMBINED PCR‐HETERODUPLEX AND PCR‐SSCP ANALYSIS FOR MATCHING OF HLA‐A, ‐B AND ‐C ALLOTYPES IN MARROW TRANSPLANTATION
Author(s) -
Pursall M.C.,
Clay T. M.,
Bidwell J.L.
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.tb00263.x
Subject(s) - heteroduplex , single strand conformation polymorphism , microbiology and biotechnology , polymerase chain reaction , typing , biology , human leukocyte antigen , dna , genetics , locus (genetics) , histocompatibility testing , antigen , gene
SUMMARY We describe a method for rapid matching of HLA‐A, ‐B and ‐C allotypes using simultaneous polymerase chain reaction‐single strand conformation polymorphism (PCR‐SSCP) and heteroduplex analysis. Electrophoresis is performed at ambient temperature without requirements for buffer cooling. SSCP and heteroduplexes are revealed as discrete spatially separated band clusters. Using HLA‐A, ‐B and ‐C locus‐specific PCR primers, matching for alleles at these loci can be performed in 5h. We tested 17 serologically matched patient‐unrelated donor pairs and found considerable microheterogeneity at the DNA level. We propose that this technology has several advantages over conventional low‐resolution typing methods and represents a potentially valuable screening method in unrelated donor selection.