Fast DNA isolation and PCR protocols for detection of methicillin-resistant staphylococci.

A rapid method that included simple boiling DNA extraction followed by a fast polymerase chain reaction (PCR) cycling protocol designed to detect mecA, which characterizes methicillin-resistant Staphylococcus aureus (MRSA), was performed. Briefly, the PCR cycling protocol consisted of pre-denaturation at 95 °C for 30 s, 30 cycles of denaturation at 94 °C for 2 s, annealing at 52 °C for 5 s, extension for 10 s, and final extension at 72 °C for 1 min. A good level of reliability of the method was verified. The study has shown that the method described here represents a rapid and accurate DNA extraction and PCR-based identification system of MRSA, thus allowing clinicians to make early identification and early implementation of control measures.