Investigating polymerase spiral reaction conditions in detecting anti-methicillin resistant gene (MecA) of Staphylococcus aureus

Abstract

     Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most dangerous pathogens on human and pets, especially as they can withstand methicillin. Identifying MRSA S. aureus strains can help with choosing proper antibiotics for treatment. The mecA gene that stipulates one of the methicillin resistant genetic combination is commonly used to detect MRSA in molecular biology techniques. The PSR (Polymerase Spiral Reaction) technique allows the amplification of the target gene sequence under isothermal conditions which is suitable for detection of MRSA. The results showed the optimal protocol of the PSR reaction at the temperature of 65 ⁰C, duration of 50 minutes with the optimal primer concentration of 1,6 μM F1+Nr/R1+N and 0,1 μM F2/R2. The detection limit of extracted DNA is 10^-3 ng/μL and the detection limit of bacterial cells is 5 bacterial cells/μL. The optimized PSR technique in this study was capable of detecting the mecA gene with a sensitivity and specificity of 100 %.