S.A. Ledwidge, B.A. Mallard1, J.P. Gibson, G.B.
Jansen and Z.H. Jiang
Anim. Genet. (2001) 32:219-221
Abstract
Multi-primer target polymerase chain reaction (MPT-PCR) is a rapid method for the identification of specific BoLA-DRB3 alleles. In a single PCR reaction, the presence of two alleles associated with increased risk, DRB3.2*23 (DRB3*2701-2703, 2705-2707) and decreased risk, DRB3.2*16 (DRB3*1501, 1502), of mastitis in Canadian Holstein can be detected. Two outer primers amplify exon 2 of DRB3. Simultaneously, two inner, allele-specific primers amplify individual alleles. Initially, 40 cows previously typed by PCR-restriction fragment length polymorphism (PCR-RFLP) were genotyped using the multi-primer approach. An additional 30 cows were first genotyped by multi-primer target PCR, then by PCR-RFLP. All animals were correctly identified and there were no false positives. This technique can readily be modified to identify other BoLA alleles of interest.
Keywords: BoLA-DRB3, bovine lymphocyte antigen, mastitis, MPT-PCR, PCR-RFLP
1Department of Pathobiology, University of Guelph