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One step PCR for detection of Staphylococcus aureus specific sequence gene and mecA gene in Northwestern Nigerian hospitals
- AS Kumurya1
© Kumurya; licensee BioMed Central Ltd. 2015
- Published: 16 June 2015
- Polymerase Chain Reaction
- Polymerase Chain Reaction Product
- Staphylococcus Aureus
- Polymerase Chain Reaction Assay
- Rapid Method
Methicillin – resistant Staphylococcus aureus (MRSA) has been noted as one of the main pathogen of public health importance. Detection of the mec A gene by polymerase chain reaction (PCR) is the gold standard for identifying methicillin-resistant Staphylococcus aureus.
In order to accelerate the procedure of identification in clinical microbiology laboratories, it is very important to havea simple and rapid method for DNA extraction. In this work, a one step PCR assay for the detection of clinically relevant antibiotic resistance gene (mec A gene) harbored by some Staphylococcus aureus isolates and for the simultaneousidentification of such isolates at the species level has been described.
In this work, a rapid method for bacterial DNA extraction directlyfrom a single colony that gave quality DNA for PCR in as littleas 15 minutes was used. PCR was used to amplify both the Staphylococcus aureus specific sequence gene and mec A gene of 100 isolates in Northwestern Nigeria with the amplicon size of 107 and 532 bp respectively. The performance and robustness of the assay was evaluated with a control strain of methicillin susceptible Staphylococcus aureus(MSSA).- ATCC 25923.
All the isolates (n=100) expressed Staphylococcus aureus specific sequence gene in their PCR products. Only 5 isolates (5.0%) were confirmed as MRSA based on the detection of mec A gene. This protocol yielded good-quality target DNA for PCRamplification. Amplifications using that DNA gave rise to goodquantities of the expected PCR fragments.
This assay offers a rapid, simple, feasible, specific, sensitive, and accurate identification of MRSA clinical isolates and could be systematically applied as a diagnostic test in clinical microbiology laboratories, facilitating the design and use of antibiotic therapy. Hence, considering that it represents a cost-effective method and helping treatment to be initiated withoutdelay.
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.