• Abdul Haque, Asma Haque, Muhammad Saeed, Aysha Azhar, Samreen Rasool, Sidra Shan, Beenish Ehsan, and Zohaib Nisar.
• Abdul Haque, PhD. Postgraduate Research Laboratory, The University of Faisalabad, Faisalabad, Pakistan.
• Pak J Med Sci. 2017 Sep 1; 33 (5): 1095-1100.

ObjectivesEmergence of methicillin resistant Staphylococcus aureus (MRSA) is a major medical problem of current era. These bacteria are resistant to most drugs and rapid diagnosis can provide a clear guideline to clinicians. They possess specific virulence factors and relevant information can be very useful. We designed this study to develop multiplex PCRs to provide rapid information.MethodsWe studied 60 Staphylococcus aureus isolates and detected methicillin resistance by cefoxitin sensitivity and targeting of mecA gene. After initial studies with uniplex PCRs we optimized two multiplex PCRs with highly reproducible results. The first multiplex PCR was developed to confirm genus, species and methicillin resistance simultaneously, and the second multiplex PCR was for screening of virulence factors.ResultsWe found 38.33% isolates as methicillin resistant. α -toxin, the major cytotoxic factor, was detected in 40% whereas β-hemolysin was found in 25% cases. Panton Valentine leucocidin was detected in 8.33% and toxic shock syndrome toxin in5% cases. The results of uniplex and multiplex PCRs were highly compatible.ConclusionsThese two multiplex PCRs when run simultaneously can provide vital information about methicillin resistance and virulence status of the isolate within a few hours as compared to several days needed by routine procedures.

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