Abortion among dairy cattle is one of the major causes of economic losses in the livestock
industry. This study describes a 1-step multiplex real-time polymerase chain reaction (PCR) to
detect Brucella spp., Leptospira spp. and Campylobacter foetus, these are significant bacteria
commonly implicated in bovine abortion. ß-actin was added to the same PCR reaction as
an internal control to detect any extraction failure or PCR inhibition. The detection limit
of multiplex real-time PCR using purified DNA from cultured organisms was set to 5 fg for
Leptospira spp. and C. foetus and to 50 fg for Brucella spp. The multiplex real-time PCR did not
produce any non-specific amplification when tested with different strains of the 3 pathogens.
This multiplex real-time PCR provides a valuable tool for diagnosis, simultaneous and rapid
detection for the 3 pathogens causing abortion in bovine.