Abstract
The need for rapid methods in order to precisely detect methicillin-resistant Staphylococcus aureus (MRSA) is extensively acknowledged. This study evaluated a quantitative real-time PCR assay targeting mecA (encoding high level resistance to methicillin) and femB (a specific genomic marker for S. aureus) genes to detect MRSA from broth culture, from serum seeded with MRSA and straight from the patient's serum. One hundred and thirty-five clinical isolates of MRSA strains and different species were utilised in this study. In addition, a pilot study with 9 patients' serum samples was performed. The sensitivity and specificity values for this assay were 99% and 100% respectively. The detection limit for this method was 1.23×102 CFU/ml from the serum seeded with MRSA cells and the limiting concentration of DNA for detection was 18 fg, which equates to 5.14 genomic DNA copies. In addition, this assay detected MRSA from patient's serum (7 out of 9) with sensitivity of 77.8%. Overall, the assay was rapid, efficient, sensitive and easy to perform.
Highlights
The purpose of this study was to investigate the effectiveness of methicillin-resistant Staphylococcus aureus (MRSA) has emerged as major cause of bloodstream infections a duplex real-time PCR assay using the TaqMan system in associated with raised risk of nosocomial infection and an detecting MRSA from serum seeded with bacteria and patients’increase in morbidity, mortality and hospitalization costs [1, 2]. sera to assess the diagnostic efficacy.Recently MRSA infections have appeared in community settings in many different countries [3, 4]
Detection limit of duplex real-time PCR The assay used in the current study was able to detect a minimum of 1.23x 102 CFU/ml from serum seeded with MRSA cells, and the limiting concentration of DNA for detection was 18 fg, which equates to 5.14 genomic DNA copies(3.5 fg/cell) [10, 11]
Detection of mecA in conjunction with femB as a second target has been reported for identification of MRSA [16, 17]
Summary
The purpose of this study was to investigate the effectiveness of MRSA has emerged as major cause of bloodstream infections a duplex real-time PCR assay using the TaqMan system in associated with raised risk of nosocomial infection and an detecting MRSA from serum seeded with bacteria and patients’increase in morbidity, mortality and hospitalization costs [1, 2]. sera to assess the diagnostic efficacy.Recently MRSA infections have appeared in community settings in many different countries [3, 4]. The purpose of this study was to investigate the effectiveness of MRSA has emerged as major cause of bloodstream infections a duplex real-time PCR assay using the TaqMan system in associated with raised risk of nosocomial infection and an detecting MRSA from serum seeded with bacteria and patients’. The sensitivity of blood cultures is markedly were taken from nine different MRSA septicemic (deep-seated reduced if blood samples are obtained during antimicrobial infection) patients to evaluate this duplex real-time PCR. Several real-time PCR methods have patients were enrolled in a clinical trial with NeuTec Pharma recently been developed for detecting MRSA directly from (Manchester, UK). Full ethical approval for analysis had been clinical samples on the same day acquired [8, 9]
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