Abstract

CARPINELLI ACEVEDO, ML et al. Multiplex PCR for simultaneous detection of mecA and pvl genes in Staphylococcus spp. Mem. Inst. Investig. Cienc. Salud [online]. 2012, vol.10, n.1, pp.5-13. ISSN 1812-9528.

The aim of this cross sectional observational study was to standardize a multiplex PCR for simultaneous detection of mecA and pvl genes in Staphylococcus spp. S. aureus strains ATCC 25923 and S.aureus ATCC 43300 were used as controls as well as an isolate of S. aureus carrying mecA and pvl genes. The boiling method was performed for DNA extraction and the detection limit was established by serial dilutions of DNA. We determined the applicability of the multiplex PCR by testing 41 isolates of S. aureus and 51 coagulase negative staphylococci (CNS) previously characterized by phenotypic methods in november 2009. The PCR products were visualized by agarose gel electrophoresis in 2% after ethidium bromide staining. The size of the amplified products of the multiplex PCR corresponded to those expected, 433bp for mecA and 533bp for pvl, with detection limits up to 0.5 ng/µL. The mecA gene was detected in 13 (31.7%) isolates of S. aureus and 29 (56.7%) CNS. The pvl gene was detected in 2 (4.9%) isolates of S. aureus and was not detected in CNS. A 100% concordance was obtained between the presence of mecA and phenotypic methods. This technique is a useful tool in the confirmation of methicillin-resistant Staphylococcus strains and identification of the pvl gene, in addition to being relatively simple, with the advantage of detecting both genes in a single reaction.

Keywords : Staphylococcus aureus; coagulase negative staphylococci; methicillin; resistance; mecA; pvl.

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