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Detection of Malaysian methicillin-resistant Staphylococcus aureus (MRSA) clinical isolates using simplex and duplex real-time PCR

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Abstract

The aim of this study was to develop a methicillin-resistant Staphylococcus aureus (MRSA) detection method based on the melting temperature analysis profiling of S. aureus clinical isolates from three different hospitals in Malaysia. Simplex and duplex real-time PCR assay was used for the simultaneous detection of nuc (species-specific) and mecA (methicillin-resistance) genes in a single SYBR Green I real-time PCR tube assay. Evaluations were based on the melting temperature (T m) analysis of the amplicons using 23 S. aureus clinical isolates including three ATCC S. aureus standard strains. Real-time PCR amplification products with melting peaks at 78.39 ± 0.4°C and 74.41 ± 0.6°C were detected for nuc and mecA genes, respectively. Each real-time PCR assay was completed within two hours. This rapid genotypic method is useful for the detection of resistant determinant (mecA) and identification of S. aureus (nuc) clinical isolates, thus benefiting patient therapy in hospitals.

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Authors and Affiliations

  1. Faculty of Agriculture and Biotechnology, Universiti Darul Iman Malaysia (UDM), Kampus Kota, Jalan Sultan Mahmud, 20400, Kuala Terengganu, Terengganu, Malaysia

    Zarizal Suhaili & Abdul Manaf Ali

  2. Antimicrobial Laboratory, Medicinal Plants Programme, Forest Biotechnology Division, Forest Research Institute Malaysia (FRIM), 52109, Kepong, Selangor, Malaysia

    Saiful Azmi Johari & Mastura Mohtar

  3. Department of Pathology, Hospital Sultanah Nur Zahirah, Jalan Sultan Mahmud, 20400, Kuala Terengganu, Terengganu, Malaysia

    Ahmad Rushdi Tan Abdullah & Affandi Ahmad

Authors
  1. Zarizal Suhaili
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  2. Saiful Azmi Johari
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  3. Mastura Mohtar
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  4. Ahmad Rushdi Tan Abdullah
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  5. Affandi Ahmad
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  6. Abdul Manaf Ali
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Correspondence to Zarizal Suhaili.

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Suhaili, Z., Johari, S.A., Mohtar, M. et al. Detection of Malaysian methicillin-resistant Staphylococcus aureus (MRSA) clinical isolates using simplex and duplex real-time PCR. World J Microbiol Biotechnol 25, 253–258 (2009). https://doi.org/10.1007/s11274-008-9887-z

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  • DOI: https://doi.org/10.1007/s11274-008-9887-z

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