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. 1995 Aug;33(8):2141–2144. doi: 10.1128/jcm.33.8.2141-2144.1995

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

V G Del Vecchio 1, J M Petroziello 1, M J Gress 1, F K McCleskey 1, G P Melcher 1, H K Crouch 1, J R Lupski 1
PMCID: PMC228351  PMID: 7559964

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.

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Selected References

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  1. Branger C., Goullet P., Boutonnier A., Fournier J. M. Correlation between esterase electrophoretic types and capsular polysaccharide types 5 and 8 among methicillin-susceptible and methicillin-resistant strains of Staphylococcus aureus. J Clin Microbiol. 1990 Jan;28(1):150–151. doi: 10.1128/jcm.28.1.150-151.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Costas M., Cookson B. D., Talsania H. G., Owen R. J. Numerical analysis of electrophoretic protein patterns of methicillin-resistant strains of Staphylococcus aureus. J Clin Microbiol. 1989 Nov;27(11):2574–2581. doi: 10.1128/jcm.27.11.2574-2581.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Goh S. H., Byrne S. K., Zhang J. L., Chow A. W. Molecular typing of Staphylococcus aureus on the basis of coagulase gene polymorphisms. J Clin Microbiol. 1992 Jul;30(7):1642–1645. doi: 10.1128/jcm.30.7.1642-1645.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hall L. M., Jordens J. Z., Wang F. Methicillin-resistant Staphylococcus aureus from China characterized by digestion of total DNA with restriction enzymes. Epidemiol Infect. 1989 Aug;103(1):183–192. doi: 10.1017/s095026880003048x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Licitra C. M., Brooks R. G., Terry P. M., Shaw K. J., Hare R. S. Use of plasmid analysis and determination of aminoglycoside-modifying enzymes to characterize isolates from an outbreak of methicillin-resistant Staphylococcus aureus. J Clin Microbiol. 1989 Nov;27(11):2535–2538. doi: 10.1128/jcm.27.11.2535-2538.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Lupski J. R., Weinstock G. M. Short, interspersed repetitive DNA sequences in prokaryotic genomes. J Bacteriol. 1992 Jul;174(14):4525–4529. doi: 10.1128/jb.174.14.4525-4529.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. OEDING P. Serological typing of Staphylococci. Acta Pathol Microbiol Scand Suppl. 1952 Jun;93:356–365. [PubMed] [Google Scholar]
  8. Prevost G., Jaulhac B., Piemont Y. DNA fingerprinting by pulsed-field gel electrophoresis is more effective than ribotyping in distinguishing among methicillin-resistant Staphylococcus aureus isolates. J Clin Microbiol. 1992 Apr;30(4):967–973. doi: 10.1128/jcm.30.4.967-973.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Rhinehart E., Shlaes D. M., Keys T. F., Serkey J., Kirkley B., Kim C., Currie-McCumber C. A., Hall G. Nosocomial clonal dissemination of methicillin-resistant Staphylococcus aureus. Elucidation by plasmid analysis. Arch Intern Med. 1987 Mar;147(3):521–524. [PubMed] [Google Scholar]
  10. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  11. Saulnier P., Bourneix C., Prévost G., Andremont A. Random amplified polymorphic DNA assay is less discriminant than pulsed-field gel electrophoresis for typing strains of methicillin-resistant Staphylococcus aureus. J Clin Microbiol. 1993 Apr;31(4):982–985. doi: 10.1128/jcm.31.4.982-985.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Sompolinsky D., Samra Z., Karakawa W. W., Vann W. F., Schneerson R., Malik Z. Encapsulation and capsular types in isolates of Staphylococcus aureus from different sources and relationship to phage types. J Clin Microbiol. 1985 Nov;22(5):828–834. doi: 10.1128/jcm.22.5.828-834.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Song M. D., Wachi M., Doi M., Ishino F., Matsuhashi M. Evolution of an inducible penicillin-target protein in methicillin-resistant Staphylococcus aureus by gene fusion. FEBS Lett. 1987 Aug 31;221(1):167–171. doi: 10.1016/0014-5793(87)80373-3. [DOI] [PubMed] [Google Scholar]
  14. Utsui Y., Yokota T. Role of an altered penicillin-binding protein in methicillin- and cephem-resistant Staphylococcus aureus. Antimicrob Agents Chemother. 1985 Sep;28(3):397–403. doi: 10.1128/aac.28.3.397. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Versalovic J., Koeuth T., Lupski J. R. Distribution of repetitive DNA sequences in eubacteria and application to fingerprinting of bacterial genomes. Nucleic Acids Res. 1991 Dec 25;19(24):6823–6831. doi: 10.1093/nar/19.24.6823. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Wenzel R., Herrmann R. Repetitive DNA sequences in Mycoplasma pneumoniae. Nucleic Acids Res. 1988 Sep 12;16(17):8337–8350. doi: 10.1093/nar/16.17.8337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Woese C. R., Maniloff J., Zablen L. B. Phylogenetic analysis of the mycoplasmas. Proc Natl Acad Sci U S A. 1980 Jan;77(1):494–498. doi: 10.1073/pnas.77.1.494. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Woods C. R., Versalovic J., Koeuth T., Lupski J. R. Whole-cell repetitive element sequence-based polymerase chain reaction allows rapid assessment of clonal relationships of bacterial isolates. J Clin Microbiol. 1993 Jul;31(7):1927–1931. doi: 10.1128/jcm.31.7.1927-1931.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. van Belkum A., Bax R., Peerbooms P., Goessens W. H., van Leeuwen N., Quint W. G. Comparison of phage typing and DNA fingerprinting by polymerase chain reaction for discrimination of methicillin-resistant Staphylococcus aureus strains. J Clin Microbiol. 1993 Apr;31(4):798–803. doi: 10.1128/jcm.31.4.798-803.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]

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