Evaluation of a duplex real-time PCR assay to detect MRSA from broth culture,human sera seeded with MRSA and from patient's serum |
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Authors: | Sawsan Awad Issam Alshami Ahmed Eid Alharbi |
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Institution: | 1.University Hospital of South Manchester NHS Foundation Trust, Wythenshawe Hospital, Southmoor Road, Manchester, M23 9LT;2.Department of Medical Microbiology and Immunology, College of Medicine, Taibah University, Almadinah, Saudi Arabia |
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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. |
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Keywords: | MRSA real-time PCR mecA femB Evaluation Duplex Sera |
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