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

Applying real-time quantitative PCR to diagnosis of freemartin in Holstein cattle by quantifyingSRYgene: a comparison experiment

“…The sterility of freemartin is explained through hormones and XX/XY chimera ( Capel & Coveney, 2004 ; Freeman, 2007 ; Gonelladiaza et al, 2012 ). In the former scenario, male hormones such as testosterone and anti-Müllerian, originally due to Wolffian development and Müllerian regression ( Loriaux, 2016 ), affect the female fetus through placental vascular anastomoses at 30–40 days of gestation and result in transformation of the general somatic habitus in female ( Jiménez, Barrionuevo & Burgos, 2013 ; Ayalavaldovinos et al, 2014 ; Refaat, Ali & Tharwat, 2015 ). The chimera scenario explains that the XY cells of a male fetus transfer to his female twin through the shared vascular anastomosis branches, and result in formation of XX/XY chimeras which hinder the development of female gonads ( Peretti et al, 2008 ).…”

“…A rapid, sensitive, and inexpensive method for identification of freemartin at birth or early age becomes an urgent need to reduce unnecessary economic losses and to preserve important hereditary material, also to avoid the delivery of detrimental genetic materials by progeny ( Biswas et al, 2015 ). Many diagnostic methods have been established for identification of freemartin, such as measurement of vaginal length ( Khan & Foley, 1994 ), blood grouping test for degree of hemolysis ( Kästli & Hall, 1978 ), karyotype analysis for XX/XY chimera ( Dunn, Johnson & Quaas, 1981 ), polymerase chain reaction (PCR) or improved PCR techniques for detection of specific fragments located on the Y chromosome ( Hirayama et al, 2007 ; Ron et al, 2010 ; Ayalavaldovinos et al, 2014 ), fluorescence in situ hybridization technique for detection of Y chromosome ( Sohn et al, 2007 ; Villagómez & Pinton, 2008 ; Rubes et al, 2009 ), quantitative detection of hormones such as progesterone, estradiol, and anti-Müllerian hormone ( Rota et al, 2002 ; Cabianca et al, 2007 ; Remnant et al, 2014 ; Kitahara et al, 2018 ), and detection of H-Y antigen qualitatively ( Wachtel et al, 1980 ). Most of the methods are based on the assumption that freemartin contains XX/XY chimera and H-Y antigen, whereas fertile heterosexual twin female does not contain XX/XY chimera or H-Y antigen.…”