Development of a multiplex PCR for the identification of Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae and Streptococcus agalactiae in milk

Abstract

In the dairy industry financial losses mainly due to loss of milk production caused by mastitis and mastitis related problems is very common. In the mastitis, identification of the infectious agent/s could be of great help. In the present study, a multiplex polymerase chain reaction (PCR) to identify five common organisms implicated in the mastitis was tried. Primers for identification of <em>Staphylococcus aureus</em>, <em>Escherichia coli </em>(<em>E. coli</em>),<em> Klebsiella pneumoniae, Pseudomonas</em> <em>aeruginosa, </em>and <em>Streptococcus agalactiae</em> were selected/ designed. DNA was extracted from the affected milk using two different methods and was subjected to multiplex PCR. It was observed from the study that a multiplex PCR to amplify four organisms<em> E. coli</em>,<em> K. pneumoniae</em>, <em>S. agalactiae </em>and <em>P. aeruginosa</em> was developed successfully to quicken the detection of common mastitis causing organisms.