African Journal of
Microbiology Research

  • Abbreviation: Afr. J. Microbiol. Res.
  • Language: English
  • ISSN: 1996-0808
  • DOI: 10.5897/AJMR
  • Start Year: 2007
  • Published Articles: 4881

Full Length Research Paper

Discriminative multiplex (hexaplex) PCR strategy for the detection of methicillin resistance and virulence factors in Staphylococcus aureus

Syed F. Hussain1, Ambrina Khatoon1, Syed M. Shahid1*, Muhammad Ismail2 and Abid Azhar1
1The Karachi Institute of Biotechnology and Genetic Engineering (KIBGE), University of Karachi, Karachi, Pakistan. 2Institute of Biomedical and Genetic Engineering (IBGE), Islamabad, Pakistan
Email: [email protected]

  •  Accepted: 19 December 2012
  •  Published: 31 January 2013

Abstract

In clinical microbiology, phenotypic characterization is laborious as well as time consuming strategy and remains less discriminative among high virulent to less virulent clinically important strains. Induction of molecular techniques, allow more accurate and quick way out for identification of Staphylococcus aureus along with its virulence capabilities. This study describes a multiplex (hexaplex) strategy for a rapid detection of methicillin resistance, simultaneously discriminating S. aureus from coagulase-negative staphylococci (CoNS) and occurrence of virulence factors. It targets the nuc (Specific forS. aureus), mec A (methicillin resistance determinant), fem A and fem B (S. aureusspecific factors essential for methicillin resistance), Luk S/F PV (encodes for Panton Valentine Leukocidin-PVL), and spa (encodes protein A). Validation of this strategy was performed using previously characterized clinical isolates of methicillin susceptible S. aureus (MSSA), methicillin resistant S. aureus (MRSA), and CoNS from different hospital facilities. Amplification results were consistent and perfectly accurate in accordance to the biochemical and resistance properties of the isolates. This molecular approach renders clinical microbiology a feasible, rapid, simple and reliable technique, discriminating MSSA, MRSA, and CoNS and provides an early and accurate way of detection. This technique will add significant contribution to prevent the wide spread dissemination of the infections as well as facilitate the designing of improved ways of treatment and cure.

 

Key words: MSSA, MRSA, mec A, panton valentine leukocidin, nuc gene, spa gene.

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