DNA typing of HLA-D/DR antigens using digoxigenin-labelled sequence specific oligonucleotide (DIG-SSO) probes, combined with polymerase chain reaction (PCR).

概要

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We amplified a DNA segment of the second exon of HLA-DRB genes with polymerase chain reaction (PCR) using pairs of DR or group specific primers, exon which codes for allogeneic determinants. Blotting the amplified products on nylon membrane filter, we tested their reactivity against dig-SSO probes without radioisotopes.As a result, DR types of DR1, DR2, DR4, DR7, DR9, DRw10, and DRw53 of 136 healthy volunteers by the DNA method were almost identical with those by the serological method that used our standard typing trays. The specificities of DRw8, DRw11, DRw12, DRw13, DRw14, and DRw52 were properly typed by the genotyping method but not, either mistyped or untyped, by the serological method. The DNA typing resolved serologically unknown HLA-DR types, and furthermore defined HLA-D specificities. Thus the DNA typing was a powerful method for detecting HLA-D/DR types.
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