AN HLA‐DR TYPING PROTOCOL USING GROUP‐SPECIFIC PCR‐AMPLIFICATION FOLLOWED BY RESTRICTION ENZYME DIGESTS

Abstract

A simple PCR‐based protocol for HLA‐DR typing suitable for a routine practice is described. The method involves, first, a PCR amplification with seven different, group‐specific (DR1, DR2, DR4, DR7, DR9, DR10, and DR3+5+6+8) primer‐pairs, and second, typing of HLA‐DR allele more exactly in DR1, DR2, DR4, and DR3+5+6+8 groups by digestion of PCR products with restriction enzymes distinguishing different HLA‐DR types within each of the groups. Altogether 24 HLA‐DR alleles, or any combination of these, can be typed. The whole procedure, starting from a blood sample, can be carried out during a single working‐day. The method was tested by typing a set of homozygous cell lines, as well as a local panel previously typed by PCR/oligotyping. Also, 227 patients waiting for transplantation were typed to test the method in a routine setting. The results suggest that this kind of approach gives reliable HLA‐DR types and works well in the routine use.