Hostname: page-component-78c5997874-4rdpn Total loading time: 0 Render date: 2024-11-04T18:21:12.577Z Has data issue: false hasContentIssue false

Restriction enzyme fingerprinting of trimethoprim resistance plasmids

Published online by Cambridge University Press:  19 October 2009

C. A. Kraft
Affiliation:
University Department of Bacteriology, Glasgoiv Royal Infirmary, Glasgow G4 0SF
M. C. Timbury
Affiliation:
University Department of Bacteriology, Glasgoiv Royal Infirmary, Glasgow G4 0SF
D. J. Platt
Affiliation:
University Department of Bacteriology, Glasgoiv Royal Infirmary, Glasgow G4 0SF
Rights & Permissions [Opens in a new window]

Summary

Core share and HTML view are not available for this content. However, as you have access to this content, a full PDF is available via the ‘Save PDF’ action button.

Restriction enzyme fingerprinting was applied to 72 transferable trimethoprim resistance plasmids to examine aspects of their epidemiology and molecular relatedness.

These plasmids had previously been divided into 25 groups according to differences in mol. wts and in antimicrobial resistance determinants. Restriction enzyme fingerprinting allowed the plasmids to be further divided into 44 different groups. The groups based on molecular weight and resistance patterns often, but not invariably, corresponded with those based on restriction enzyme fingerprints. Some plasmids with the same mol. wt and resistance pattern had different digest fingerprints and conversely, although more rarely, plasmids which differed in molecular weight by as much as 10 MDa or in resistance pattern by one resistance marker, had indistinguishable fingerprints.

The plasmids were initially divided into three broad categories according to which restriction enzymes gave fingerprints of 6–20 fragments. These categories differed in the molecular weights of the plasmids contained, the numbers of resistance markers, and the proportions of the plasmids which carried transposon Tn7.

Some plasmids were more widespread and persistent than others with the same mol. wt and resistance pattern but with a different restriction enzyme fingerprint.

Thus, application of this technique has shown the trimethoprim resistance plasmids studied to be more diverse than was indicated by determination of mol. wt and resistance pattern, and has indicated changes in the plasmid pool over the 3 years during which they were collected.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1987

References

REFERENCES

Beul, H. A., Eichel-Streiber, Ch. von, Schreiner, M., Schwindling, F. P., Weinblum, D., Zollner, E. J. & Dierich, M. (1985). Characterisation of cryptic plasmids in clinical isolates of Bacleroides fragilia. Journal of Medical Microbiology 20, 3948.CrossRefGoogle Scholar
Birnboim, H. C. & Doly, J. (1979). A rapid alkaline extraction procedure for screening recombinant DNA. Nucleic Acids Research 7, 15131523.CrossRefGoogle Scholar
Datta, N. (1977). R-factors in enterobacteriaceae. In R-factor, Drug Resistance Plasmid (ed. Mitsuhashi, S.) pp. 255272. Tokyo: Tokyo University Press.Google Scholar
Fleming, M. P., Datta, N. & Gruneberg, R. N. (1972). Trimethoprim resistance determined by R-factors. British Medical Journal 1, 726728.CrossRefGoogle ScholarPubMed
Ish-Horowitz, D. & Burke, J. F. (1981). Rapid and efficient cosmid cloning. Nucleic Acids Research 9, 29892998.CrossRefGoogle Scholar
Kraft, C. A., Platt, D. J. & Timbury, M. C. (1983). Distribution and transferability of plasmids in trimethoprim resistant urinary Escherichia coli. Journal of Medical Microbiology 16, 433441.CrossRefGoogle ScholarPubMed
Kraft, C. A., Platt, D. J. & Timbury, M. C. (1984). Distribution and transferability of plasmids in trimethoprim-resistant urinary strains of Escherichia coli: a comparative study of hospital isolates. Journal of Medical Microbiology 18, 95105.CrossRefGoogle ScholarPubMed
Kraft, C. A., Platt, D. J. & Timbury, M. C. (1985). Trimethoprim resistance in urinary coliforms from patients in the community: plasmids and R-transfer. Journal of Antimicrobial Chemotherapy 15, 311317.CrossRefGoogle ScholarPubMed
Kraft, C. A., Timbury, M. C. & Platt, D. J. (1986). Distribution and genetic location of Tn7 in trimethoprim resistant E. Coli. Journal of Medical Microbiology 22, 125131.CrossRefGoogle Scholar
Lyon, B. R., Iuorio, J. L., May, J. W. & Skurray, R. A. (1984). Molecular epidemiology of multi-resistant Staphylococcus aureus in Australian hospitals. Journal of Medical Microbiology 17, 7989.CrossRefGoogle Scholar
O'Brien, T. F., Hopkins, J. D., Gilleece, E. S., Madeiros, A. A., Kent, R. K., Blackburn, B. O., Holmes, M. B., Reardon, J. P., Vergeront, J. M., Schell, W. L., Chirstenson, E., Bissert, M. L. & Morse, E. V. (1982). Molecular epidemiology of antibiotic resistance in Salmonella from animals and human beings in the United States. New England Journal of Medicine 307, 16.CrossRefGoogle ScholarPubMed
Platt, D. J., Chesham, J. S. & Kristinsson, K. G. (1986). R-transfer in vivo: a prospective study. Journal of Medical Microbiology 21, 325330.CrossRefGoogle ScholarPubMed
Platt, D. J., Chesham, J. S., Brown, D. J., Kraft, C. A. & Taggart, J. (1986). Restriction enzyme fingerprinting of enterobacterial plasmids: a simple strategy with wide application. Journal of Hygiene 97, 205210.CrossRefGoogle ScholarPubMed
Richards, H. & Nugent, M. (1979). The incidence and spread of Tn7. In Plasmids of Medical, Environmental and Commercial Importance (ed. Timmis, K. N. & Puhler, A..) Elsevier/North Holland Biomedical Press.Google Scholar
Rubens, C. E., Farrar, W. E. Jr, McGee, X. A. & Schaffner, W. (1981). Evolution of a plasmid mediating resistance to multiple antimicrobial agents during a prolonged epidemic of nosocomial infections. The Journal of Infectious Diseases 143, 170181.CrossRefGoogle ScholarPubMed