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Title

Evaluation of a duplex real-time PCR assay to detect MRSA from broth culture, human sera seeded with MRSA and from patientís serum

 

Authors

Sawsan Awad1, Issam Alshami2 & Ahmed Eid Alharbi2*

 

Affiliation

1University Hospital of South Manchester NHS Foundation Trust, Wythenshawe Hospital, Southmoor Road, Manchester, M23 9LT; 2Department of Medical Microbiology and Immunology, College of Medicine, Taibah University, Almadinah, Saudi Arabia

 

Email

ishami@taibahu.edu.sa; *Corresponding author

 

Article Type

Hypothesis

 

Date

Received October 01, 2013; Accepted October 03, 2013; Published November 11, 2013

 

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 patientís 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 patientsí serum samples was performed. The sensitivity and specificity values for this assay were 99% and 100% respectively. The detection limit for this method was 1.23x102 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 patientís serum (7 out of 9) with sensitivity of 77.8%. Overall, the assay was rapid, efficient, sensitive and easy to perform. 

 

Keywords

MRSA, real-time PCR, mecA, femB, Evaluation, Duplex, Sera.

 

Citation

Awad et al. Bioinformation 9(18): 896-900 (2013)

 

Edited by

P Kangueane

 

ISSN

0973-2063

 

Publisher

Biomedical Informatics

 

License

This is an Open Access article which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. This is distributed under the terms of the Creative Commons Attribution License.