Development and application of a triplex‐PCR assay for rapid detection of methicillin‐resistant Staphylococcus aureus from pigs

A triplex‐PCR assay was developed and evaluated for rapid detection of methicillin‐resistant Staphylococcus aureus (MRSA) recovered from various biological samples of pig. Three sets of primers were designed to target mecA, 16S rRNA and nuc genes of MRSA. The specific amplification generated three bands on agarose gel, with sizes 280 bp for mecA, 654 bp for 16S rRNA and 481 bp for nuc, respectively. A potential advantage of the PCR assay is its sensitivity with a detection limit of 102 CFU per ml of bacteria. In all, 79 MRSA isolates recovered from various samples of pigs were subjected to the amplification by the triplex‐PCR assay and all the isolates yielded three bands corresponding to the three genes under this study. No false‐positive amplification was observed, indicating the high specificity of the developed triplex‐PCR assay. This assay will be a useful and powerful method for differentiation of MRSA from methicillin‐sensitive S. aureus, coagulase‐negative methicillin‐resistant staphylococci and coagulase‐negative methicillin‐sensitive staphylococci. Significance and Impact of the Study: The study reports the development and evaluation of a triplex‐PCR assay for the rapid detection of methicillin‐resistant Staphylococcus aureus (MRSA) from pigs. There is an urgent public health need for early and reliable detection of MRSA infection not only to reduce the cost of treatment but also to direct appropriate antibiotic therapy for its better management. This assay will be very useful for the rapid detection of MRSA from pigs.