Researches Concerning the Molecular Testing of Basset Hound Dogs For Detecting PMDS

Abstract

The Persistent MA¼llerian Duct Syndrome (PMDS), a rare form of dog male pseudo-hermaphroditism, it has been previously studied from a molecular aspect in miniature schnauzers by Matsuu et al. (2009), which detected an affected case, positive to Sry gene detection by PCR. The molecular testing for PMDS syndrome has not yet been performed at Basset Hound dogs, although affected cases exist. The aim of our research was to test the previously published protocol for Sry gene detection in an affected PMDS Basset Hound family so as to develop a possible molecular method for genotyping. The study was performed on 11 Basset Hound individuals, among which 1 affected male and 2 non-affected carrier. The protocol was applied accordingly to the one previously published by Matsuu et al. (2009). The DNA was extracted from peripheral blood samples using a specific DNA extraction kit (Isolate II, Bioline, England). Briefly, the PCR technique was performed on 25 µl of mixture containing: 12.5 µl MasterMix (Bioline), 1 µl Primer Forward and 1 µl of Primer Reverse, 6.5 µl DNA-free nuclease PCR water (Sigma Aldrich, Germany). The PCR products were visualized in 2% agarose gel stained with EvaGreen (Sigma, England). The tested protocol has shown specificity in the case of the positive affected male, showing a ligh-band at 104 base pairs in the agarose gel. All the other samples were tested negative for Sry gene. Given the fact that this protocol does not use a restriction enzyme for possible detection of genotypes and carriers, we could not differentiate the carriers from the non-affected individuals. This protocol can be used only in the case of affected males but does not distinguish the carriers from the non-carriers. We should furtherly investigate other candidate genes for PMDS, such as MISRII, for the possible detection of female and male carriers.