Multiplex polymerase chain reaction for simultaneous detection of major mastitis-causing pathogens in buffalo milk

Abstract

Mastitis is one of the major diseases in the dairy sector and to efficiently treat and minimize its effect on dairy industry, a sensitive, rapid, and specific test for identifying mastitis-causing pathogens is required. PCR-based method provides a promising option, comparing with bacteriological method. Improved PCR method was applied to simultaneously detect various pathogens in one assay. Here, the possible use of the multiplex PCR , capable of detecting simultaneously four of the most common mastitis-causing pathogens in milk including Staphylococcus aureus, Streptococcus agalactiae, Streptococcus dysgalactiae and Streptococcus uberis were tested using DNA extracted from buffalo milk. Twenty five milk samples tested using mPCR were evaluated on the level of severity through California Mastitis Test . Sensitivity of mPCR was assessed by comparing the results from bacteriological culture. Results from both methods matched in 56% of samples; while 44% were detected positive in mPCR but had no growth in the culture. This study suggests that mPCR can be a practical tool applicable in dairy laboratories for rapid detection of mastitis-causing pathogens in milk that would make significant improvements in the control and prevention of mastitis.