title: 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.
authors.affiliation: Molecular Biology Institute, University of Scranton, Pennsylvania 18510, USA.
authors.forename: V G
authors.initials: VG
authors.lastname: Del Vecchio
authors.forename: J M
authors.initials: JM
authors.lastname: Petroziello
authors.forename: M J
authors.initials: MJ
authors.lastname: Gress
authors.forename: F K
authors.initials: FK
authors.lastname: McCleskey
authors.forename: G P
authors.initials: GP
authors.lastname: Melcher
authors.forename: H K
authors.initials: HK
authors.lastname: Crouch
authors.forename: J R
authors.initials: JR
authors.lastname: Lupski
mesh_terms: D015373:Bacterial Typing Techniques; D001483:Base Sequence; D003428:Cross Infection / Q000382:microbiology; D016172:DNA Fingerprinting / Q000379:methods; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005069:Evaluation Studies as Topic; D005456:Fluorescent Dyes; D005838:Genotype; D006801:Humans; D016106:Methicillin Resistance / Q000235:genetics*; D017720:Molecular Epidemiology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D012091:Repetitive Sequences, Nucleic Acid; D015203:Reproducibility of Results; D013203:Staphylococcal Infections / Q000382:microbiology; D013211:Staphylococcus aureus / Q000145:classification / Q000187:drug effects* / Q000235:genetics*
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D005456:Fluorescent Dyes