SYBR Green I-based Real-time PCR Assay and Melting Curve Analysis for Rapid Detection of Staphylococcus aureus from Raw Milks Samples
2008
Jung, J.H. (Sahmyook University, Seoul, Republic of Korea) | Jeong, S.Y. (Sahmyook University, Seoul, Republic of Korea) | Lee, S.J. (Sahmyook University, Seoul, Republic of Korea) | Choi, S.S. (Sahmyook University, Seoul, Republic of Korea), E-mail: [email protected]
The aim of this study was to develop a LightCycler-based real time PCR (LC-PCR) assay and to evaluate its diagnostic use for the detection of Staphylococcus aureus in raw milk samples. Following amplification of 113 bp of coa gene encoding an coagulase precursor specific for Staphylococcus aureus, melting curve and DNA sequencing analysis was performed to verify the specificity of the PCR products. Amplification of 209 bp gene encoding an altered penicillin-binding protein, PBP2a (mecA), melting curve analysis and DNA sequencing analysis was performed to verify methicillin resistance Staphylococcus aureus (MRSA). According to this study, 6 of 647 raw milk samples showed S. aureus positive and 2 of them showed a mecA positive and the detection limit was 10 fg of DNA. And we also isolated Staphylococcus chromogenes a causative agent of exudative epidermitis in pigs and cattle from 3 samples.
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