COMBINED PCR‐HETERODUPLEX AND PCR‐SSCP ANALYSIS FOR MATCHING OF HLA‐A, ‐B AND ‐C ALLOTYPES IN MARROW TRANSPLANTATION
作者:
M.C. Pursall,
T. M. Clay,
J.L. Bidwell,
期刊:
International Journal of Immunogenetics
(WILEY Available online 1996)
卷期:
Volume 23,
issue 1
页码: 41-53
ISSN:1744-3121
年代: 1996
DOI:10.1111/j.1744-313X.1996.tb00263.x
出版商: Blackwell Publishing Ltd
数据来源: WILEY
摘要:
SUMMARYWe 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. UsingHLA‐A, ‐Band ‐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 unre
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