A Rapid Multiplex PCR Method for the Diagnose of Freemartin Syndrome in Domestic Cattle (Bos taurus)

Abstract

The majority of heifer calves born as hetero–sexual twin births are sterile freemartins. It is Revised: 2014–01–09 important that this condition be diagnosed at an early age because freemartin heifers can’t be Accepted: 2014–01–09 used as replacement stock. Different methods are available for the diagnosis of freemartinism, however molecular methods are preferred due to their accuracy and shorter duration of process. In the present study, freemartinism status of 40 female calves of heterosexual Key Words: Freemartinism, multiple births was investigated by multiplex polymerase chain reaction (multiplex PCR). Bovine, XX/XY Chimerism, The reaction was carried out using two primer sets for the Sry and K–casein genes of Multiplex PCR domestic cattle. The PCR product was analyzed by agarose gel (3%) electrophoresis, which allowed to identify the genotypes of the animals studied: normal males (453 bp and 163 bp), normal females (453 bp), and 39 (97.50%) study cases that showed chromosome chimerism (freemartin, 453 bp and 163 bp). Compared to the polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) technique, this method (i.e., multiplex PCR) proved a far cheaper and quicker (approximately 27 minutes) way to diagnose freemartinism.