HLA class I typing by sequence specific oligonucleotide probes after genomic DNA amplification

HLA class I typing by sequence specific oligonucleotide probes after genomic DNA amplification

Abstracts 31 #45 4.2 #46 HLA CLASS I TYPING BY SEQUENCE SPECiFiCOLIGONUCLEOTIDEPROBES AFTER GENOMIC DNA AMPLIFICATION. ~ Z Jin, R Khan, B Dupont a...

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Abstracts

31

#45 4.2

#46

HLA CLASS I TYPING BY SEQUENCE SPECiFiCOLIGONUCLEOTIDEPROBES AFTER GENOMIC DNA AMPLIFICATION. ~ Z Jin, R Khan, B Dupont and SY Yang, Laboratory of Immunogenetics, Memorial S~an-Kettering Cancer Center, New York, NY. Gene specific amplification utilizing polymerase chain reaction {PCR} combined with sequence specific oligonuclaotide probes {SSOPs} has been successfully applied for the purpose of determining the HLA typing of class II alleles. The same PCR-based oligonucleotide typing was developed for HLA class I alleles" using the HLA-A locus as a model. The HLA-A locus specific primers, which flank exon 2 and exon 3, and SSOPs were designed besed on presently available class I sequences. Panel cel|s of over 100 unrelated individuals, mostly heterozygous for HLA-A, which had been HLA typed by serology and one dimensional isoelectric focusing {1D-IEFL were selacted to include all WHO designated HLA-A specificitlas as well as IEF subtypes. When 37 SSOPs were tested on the cell panel, three SSOPs, aH of the serologically defined HLA-A alleles could be identified by their unique hybridization patterns. In addition, five A2, three A26, and two A30 subtypes detected by 1D-IEF could also be ident!fied by this penel of SSOPs. Although many of the SSOP-definabla alleles have not been characterized by nuclaotide sequences, a unique hybridization pattern for each aUetehas made typing possible. The HLA-A related HLA-H, previously known as HLA-AR, and other class I pseudogenes did not interfere with the typing for HLA-A allalas. Hybridization patterns for HLA-H locus-specific SSOPs suggest that not al| haplotypes carry this gene.

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