Development and application of a triplex-PCR assay for rapid detection of methicillin-resistant Staphylococcus aureus from pigs |
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| Authors: | S Rajkhowa SR Pegu GP Patil RK Agrawal |
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| Institution: | 1. Animal Health Laboratory, ICAR-National Research Centre on Pig, Rani, Guwahati, Assam, India;2. ICAR- National Research Centre on Meat, Boduppal post, Chengicherla, Hyderabad, India;3. Indian Veterinary Research Institute, Izatnagar, UP, India |
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| Abstract: | 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. |
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| Keywords: | methicillin-resistant Staphylococcus aureus pig sensitivity specificity triplex PCR |
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