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Substitution of Human for Horse Urine Disproves an Accusation of Doping

NCJ Number
Journal of Forensic Sciences Volume: 53 Issue: 5 Dated: September 2008 Pages: 1145-1148
Silvina Diaz Ph.D.; Mariana E. Kienast Ph.D.; Egle E. Villegas-Castanasso Ph.D.; Natalia L. Pena; Marcos M. Manganare; Diego Posik Ph.D.; Pilar Peral-Garcia Ph.D.; Guillermo Giovambattista Ph.D.
Date Published
September 2008
4 pages
In order to detect the switching and/or manipulation of samples, the owner of a stallion who was suspected of doping his horse asked the authors’ lab to perform a DNA test on a positive doping urine sample, in order to compare the urine DNA profile with blood and hair DNA profiles from the same stallion.
The study found that only blood and hair samples produced results in the 12 micro satellites used for horses. Identical genetic profiles were evident in blood and hair; however, the urine sample did not produce any results, even though positive controls were seen. Several explanations for negative results were possible, including low DNA quality in the urine sample or the presence of inhibitors of the PCR reaction in the template solution. Another explanation considered was that the species origin of the urine was other than horse. In order to test the last possibility, the mitochondrial cytochrome b (Cyt b) was amplified in order to determine the species origin of the urine sample. The band patterns obtained from the urine sample resembled those from the human DNA control. Confirmation testing showed that the alleged horse urine was in fact of human origin, indicating the manipulation of samples. This evidence supported the breeder’s contention that he was innocent of suspected doping of the stallion. Descriptions of material and methods address the animal samples, DNA isolation, horse DNA typing, species-specific determination by RFLP analysis of PCR products, and DNA sequencing and Sequence analysis. 2 figures and 22 references