Molecular genotyping of methicillin-resistant Staphylococcus aureus via fluorophore-enhanced repetitive-sequence PCR

Abstract

Methicillin resistance in Staphylococcus aureus is a frequent cause of nosocomial and community-acquired infections. Accurate, rapid epidemiologic typing is crucial to the identification of the source and spread of infectious disease and could provide detailed information on the generation of methicillin-resistant S. aureus (MRSA) strains. The high degree of genetic relatedness of MRSA strains has precluded the use of more conventional methods of genetic fingerprinting. A rapid DNA fingerprinting method that exploits PCR amplification from a DNA repeat sequence in MRSA is described. The random chromosomal distribution of this repeat sequence provides an ideal target for detecting DNA fragment patterns specific to individual MRSA strains. Two PCR fingerprinting methods which use an oligonucleotide primer based on a repetitive sequence found in Mycoplasma pneumoniae are presented. The repetitive element sequence-based PCR (rep-PCR) and fluorophore-enhanced rep-PCR (FERP) can identify epidemic strains among background MRSA. The combination of oligonucleotide primers labeled with different fluorescent dyes allowed simultaneous FERP fingerprinting and mecA gene detection. Eight different fingerprint patterns were observed in MRSA strains collected from different sources. These techniques provide a rapid discriminatory means of molecular epidemiologic typing of MRSA involved in nosocomial infections.

Cationic compounds with activity against multidrug-resistant bacteria: interest of a new compound compared with two older antiseptics, hexamidine and chlorhexidine

“…Antimicrobial resistances were determined by the automated instrument VITEK2 (bioMerieux, Marcy L'Etoile, France). The presence of resistance genes was investigated by PCR multiplex analysis adapted from methods previously described by Dutka-Malen et al [10] for van genes, or Del Vecchio et al [11] for mecA genes. Strains were grown on Mueller Hinton agar (BD, 225250) or in Mueller Hinton broth (MHB) (BD, 275730), complemented with 5% lysed sheep blood for the streptococci.…”

REP-PCR tracking of the origin and spread of airborne Staphylococcus aureus in and around chicken house

“…It has been shown that this method is reproducible and accurate. Vecchio et al (1995) showed that their PCR products were highly reproducible and stable when trying to distinguish 170 stains of Mycoplasma pneumoniae using RW3A of RepMP3 as primer. Van der Zee et al (1999) showed that REP-PCR results were similar to pulse field gel electrophoresis (PFGE) analysis.…”

Decolonization of methicillin-resistant Staphylococcus aureus using oral vancomycin and topical mupirocin

“…Methicillin resistance was determined by oxacillin disk diffusion test and by oxacillin E test [24,28]. Isolates were confirmed to be methicillin resistant by polymerase chain reaction (PCR) for the mecA gene [29]. Phage typing was performed at the National Institute of Public Health and the Environment.…”