Rapid detection of mecA in methicillin resistant Staphylococcus aureus using Cycling Probe Technology

A novel method has been developed for the detection of the mecA gene, that confers the principle mechanism of methicillin resistance in staphylococci. Cycling Probe Technology (CPT) is a rapid, simple, isothermal method for t he detection of specific target sequences. CPT utilizes a unique chimeric D NA-RNA-DNA probe sequence that provides an RNase H sensitive scissile I ink when hybridized to a complementary target DNA sequence. In the presence of target DNA, the cycling reaction converts full-length chimeric probe into cleaved probe fragments, which accumulate and are quantified. A cycling pro be designed for detection of a specific sequence within the mecA gene was u sed to develop a culture confirmation assay for methicillin resistant Staph ylococcus aureus. The CPT assay was used to screen 238 S. aureus isolates a nd the results were in complete agreement with detection of the mecA gene b y polymerase chain reaction (PCR). Detection of mecA should be considered t he gold standard for determining methicillin resistance in S. aureus. This study demonstrates the feasibility of using CPT to meet this need. (C) 1999 Academic Press.