Tm. Limm et al., HLA-DQA1 ALLELE AND SUBALLELE TYPING USING NONCODING SEQUENCE POLYMORPHISMS - APPLICATION TO 4AOHW CELL PANEL TYPING, Human immunology, 38(1), 1993, pp. 57-68
HLA-DQA1 typing of the 4AOHW cell panel is presented using a novel str
ategy that exploits both intron and exon polymorphisms. Intron sequenc
es adjacent to the variable HLA-DQA1 second exon exhibit stable polymo
rphisms that are specific for locus alleles and certain suballelic DR/
DQ haplotypes. A PCR-RFLP method has been developed that is based on a
mplification of a: 780-bp segment extending from intron I through exon
2 to intron 2. Stable sequence polymorphisms provide restriction enzy
me sites and confer mobility variations detected on polyacrylamide min
igel electrophoresis. Direct band comparison of amplified products and
restriction fragments with known standards facilitates pattern compar
ison, obviating the requirement for accurate molecular weight determin
ation. This method, using only two enzymes, identifies a total of 11 a
llelic and suballelic groups, including all eight DQA1 alleles encoded
at the second exon.