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Development of a PCR assay for identification of antibiotic resistance determinants at Staphylococcus aureus

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Author(s): Cornelia CRĂCIUNAŞ | Anca BUTIUC-KEUL | Mirela FLONTA | Ariana ALMAŞ | Andreea BRAD | Marta SIGARTEU

Journal: Analele Universitatii din Oradea, Fascicula Biologie
ISSN 1224-5119

Volume: TOM XVII;
Issue: 2;
Start page: 248;
Date: 2010;
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Keywords: Staphylococcus aureus | antibiotic resistance | PCR

ABSTRACT
The aim of the present study was to develop a PCR assay for identification of antibiotic resistance genes at Staphylococcus aureus clinical isolates. Thirty five isolates from the Emergency County Hospital and Clinical Hospital for Infectious Diseases in Cluj-Napoca, Romania, were analyzed. The bacteria were isolated over two years period, in 2007 and 2009 and serotyped at the Clinical Hospital of Infectious Disease, Cluj-Napoca. In order to develop a PCR assay for identification of antibiotic resistance genes at S. aureus clinical isolates, the bacterial strains were tested for the presence of the tst, sea, mecA, femA, ermC and ermA genes using the polymerase chain reaction and gene-specific primers. We succeeded in amplifying mecA gene to all isolates that exhibit methicillin resistance. Our result concord 100% with standard culture method but they were obtained in 5 hours instead of four days. In order to increase the efficiency of PCR typing and reduce reagent costs, multiple sets of primers were included in a single reaction tube in a multiplex PCR. We succeeded in elaborating a multiplex PCR for mecA and femA genes.
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