Triplex PCR combined with magnetic separation strategy for rapid and specific detection of methicillin-resistant Staphylococcus aureus in hospital samples

Abstract Due to the massive use of antibiotics, clinical highly pathogenic bacteria methicillin-resistant Staphylococcus aureus (MRSA) have emerged. Given its non-uniform drug resistance, it brings numerous difficulties for clinical diagnostic tests. This study aimed to establish a fast and accurate method to detect MRSA in hospital samples. In order to shorten the detection time and eliminate the influence of matrix on the detection results, so as to improve the detection sensitivity. In this study, magnetic separation (MS) technology was used for pre-treatment of samples, and triplex PCR was established using mecA, nuc and flic genes to realize the detection of MRSA. The results showed that the lowest detection limit of MRSA in blood samples and cerebrospinal fluid could reach 102 CFU/mL and 103 CFU/mL. Therefore, the MS technology combined with triplex PCR assay established in this study is expected to provide a strong basis for the detection of MRSA in hospital samples.