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KMID : 0876920040080020023
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Kwandong Medical Journal
2004 Volume.8 No. 2 p.23 ~ p.32
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Multiplex PCR for Rapid Detection of Methicillin-Resistant Staphylococcus aureus
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Park Yoon-Sun
Shin Woon-Seob
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Abstract
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Methicillin-resistant Staphylococcus aureus (MRSA) is a common cause of nosocomial infections as well as community acquired infections. The rapid identification of MRSA is very important for the appropriate antibiotic therapy, and MRSA must be differentiated from coagulase-negative staphylococci, which may be members of normal microbial flora. However, the conventional phenotypic methods for MRSA detection often lack sensitivity, because of the hetrogeneous expression of methicillin resistance and the dependence of resistance expression on the test conditions. In this study, we developed a multiplex PCR (MPCR) assay for rapid and accurate detection of MRSA. Two pairs of primers were used, which yielded two PCR products. One PCR product is a 310-bp mecA-based PCR fragment specific for methicllin resistance, and the other is a 108-bp S. aureus-specific DNA fragment. The multiplex PCR developed in this study showed excellent specificity and sensitivity for MRSA detection. When three methods for DNA extraction from MRSA were compared, lysostaphin methods and bead beating methods were more efficient than the heating methods. Therefore, the MPCR using two pairs of primers is thought to be a very sensitive and specific tool for rapid detection of MRSA, especially when the DNA of MRSA is extracted by bead beating method.
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KEYWORD
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MRSA, MPCR, mecA S. aureus-specific DNA
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