Multi-primer target PCR for rapid identification of bovine DRB3 alleles.
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.
Ledwidge
S A
SA
Center for the Genetic Improvement of Livestock, Department of Animal and Poultry Science, University of Guelph, Guelph, Ontario, Canada N1G 2W1. sledwidg@uoguelph.ca
Mallard
B A
BA
Gibson
J P
JP
Jansen
G B
GB
Jiang
Z H
ZH
D000483:Alleles; D000818:Animals; D002417:Cattle / Q000235:genetics* / Q000276:immunology; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D005260:Female; D005838:Genotype; D000949:Histocompatibility Antigens Class II / Q000032:analysis / Q000235:genetics* / Q000276:immunology*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*
C423282:BoLA-DRB3 antigen; D000949:Histocompatibility Antigens Class II; D004247:DNA