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Real Time PCR technique applied to the study of intersexed animals (bovines and canines)

Abstract:

The reproductive disorders in domestic animals often are associated with chromosomal abnormalities. In recent years, the study of chromosomal complement of intersexed animals (ie. presence of Y sex chromosome in female-like animals) was supplemented with the development of molecular techniques, allowing the realization of more precise analysis. The objective of this work was the development of the technique of real-time PCR to detect specific sequences of the Y sex chromosome. For this, we studied two bovine freemartin females and one case of intersex in canine. Some specimens of normal males and females as negative and positive controls respectively were used. The DNA was extracted from peripheral blood by conventional phenolchloroform technique. DNA samples were analyzed for the SRY gene sequences (canine) and a repeated specific sequence of the Y chromosome bovine (BRY4). The amplification of the sequences in real time thermocycler was performed using Biotools QuantyMix Corbett and SYG Kit. A characteristic fluorescence curve was obtained for each sequence, consistent with the positive controls used, Y chromosome sequences were identified in intersexed animals (female freemartin and canine). It is shown a fast, simple and highly specific for the diagnosis of Y chromosome sequences in intersexed animals.