title: Comparison of cytogenetics and polymerase chain reaction based detection of the amelogenin gene polymorphism for the diagnosis of freemartinism in cattle.
abstract: A polymerase chain reaction (PCR) assay which detects a sex-based polymorphism in the bovine amelogenin locus was modified and compared to conventional cytogenetic analysis for diagnosis of freemartinism (XX/XY chimerism) in cattle. The PCR assay is more sensitive than cytogenetic analysis for detection of XY cells, with the limit of detection of the assay falling between 0.2% and 1% XY cells. Seventy-three heifer blood samples submitted for evaluation of freemartinism to the University of Minnesota Diagnostic Laboratory were tested using both cytogenetic and PCR techniques. Poor-quality samples precluded successful lymphocyte culture and recovery of mitotic nuclei for cytogenetic evaluation in 17 cases (23%). Two of these samples (2.7%) also failed to amplify with PCR. There was 100% agreement in the results from the 56 samples that were suitable for testing using both techniques. This PCR-based assay provides an alternative to the more laborious cytogenetic evaluation for diagnosis of freemartinism.
authors.affiliation: Department of Veterinary Clinical Sciences and Veterinary Diagnostic Laboratory, College of Veterinary Medicine, University of Minnesota, 1352 Boyd Avenue, Saint Paul, MN 55108, USA. mcnie001@umn.edu
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: McNiel
authors.forename: Nicole J
authors.initials: NJ
authors.lastname: Madrill
authors.forename: Amy E
authors.initials: AE
authors.lastname: Treeful
authors.forename: Lance C
authors.initials: LC
authors.lastname: Buoen
authors.forename: Alvin F
authors.initials: AF
authors.lastname: Weber
mesh_terms: D053523:Amelogenin; D000818:Animals; D002417:Cattle; D020732:Cytogenetic Analysis / Q000662:veterinary; D004247:DNA / Q000737:chemistry / Q000235:genetics; D003746:Dental Enamel Proteins / Q000235:genetics*; D004587:Electrophoresis, Agar Gel / Q000662:veterinary; D005260:Female; D005611:Freemartinism / Q000175:diagnosis* / Q000235:genetics*; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D011110:Polymorphism, Genetic; D012680:Sensitivity and Specificity; D014960:X Chromosome / Q000235:genetics; D014998:Y Chromosome / Q000235:genetics
chemical_list: D053523:Amelogenin; D003746:Dental Enamel Proteins; D004247:DNA