Rapid Detection of Staphylococcus aureus and Methicillin-Resistant Staphylococcus aureus by Multiplex Real-Time Fluorescent PCR.

Abstract

Staphylococcus aureus (S.aureus) is a common Gram-positive human pathogen that causes community-acquired and nosocomial infections, with diverse clinical manifestations ranging from local superficial lesions and food poisoning to fatal systemic infections. The discovery of antibiotics significantly reduced the mortality rate, but the problem of drug resistance has since become increasingly prominent. Since the first identification of MRSA in 1960, this strain has emerged as a global public health threat. MRSA is a major pathogen of nosocomial infections, capable of causing various severe diseases such as endocarditis, chronic osteomyelitis, pneumonia, pyogenic arthritis, and bacteremia. Therefore, rapid and accurate detection of S. aureus and its drug resistance is crucial for guiding clinical treatment. Current routine detection methods for S.aureus and MRSA have significant limitations. The traditional bacterial culture method, serving as the "gold standard" for decades, can provide definitive species identification and drug sensitivity results, but the process is time-consuming, taking 48 to 72 h. Furthermore, this method is susceptible to contamination and relies on specialized laboratory facilities and skilled technicians. Serological testing achieves non-invasive diagnosis by detecting S.aureus antibodies in patient serum, but it cannot distinguish between active infections and past infections, nor can it identify drug-resistant strains (such as MRSA). This study focuses on developing a novel multiplex real-time fluorescence PCR detection method to overcome the above-mentioned limitations. This method designs specific primers and TaqMan fluorescent probes targeting the species-specific nuc gene of S.aureus and the mecA gene mediating methicillin resistance, enabling simultaneous amplification and detection of S.aureus and MRSA in a single reaction system. This technology greatly reduces the detection time, providing a rapid, accurate, and cost-effective solution for S.aureus and MRSA detection. This innovative approach greatly improves clinical diagnostic efficiency and facilitates the early implementation of targeted antibiotic therapy, making important contributions to controlling drug-resistant bacterial infections.