A FAST AND HIGHLY SENSITIVE METHOD FOR DETECTING FREEMARTINISM IN BOVINE TWINS USING IMMUNOMAGNETIC BEADS AND Y-SPECIFIC PCR PRIMERS

A method for detecting male cells in the blood of the female calf in b ovine heterosexual twin pregnancies has been established. Nucleated ce lls were isolated from full blood by immunomagnetic separation, lysed by boiling and then subjected to polymerase chain reaction (PCR) ampli fication with Y chromosome specific primers. Diagnosis was achieved wi thin one day. The method was successfully used on blood samples that h ad been stored at +4-degrees-C for more than one month. Dilution of ma le blood in female blood showed that XY cells were detectable down to a concentration of 0.1%. This method should be amenable to automatizat ion and can be adapted to any PCR-based genetic test.