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Restriction enzyme fingerprinting of enterobacterial plasmids: a simple strategy with wide application

Published online by Cambridge University Press:  19 October 2009

D. J. Platt ,
J. S. Chesham ,
D. J. Brown ,
C. A. Kraft  and
J. Taggart
D. J. Platt
Affiliation:
University Department of Bacteriology, Glasgow Royal Infirmary, Castle Street, Glasgow G4 0SF
J. S. Chesham
Affiliation:
University Department of Bacteriology, Glasgow Royal Infirmary, Castle Street, Glasgow G4 0SF
D. J. Brown
Affiliation:
University Department of Bacteriology, Glasgow Royal Infirmary, Castle Street, Glasgow G4 0SF
C. A. Kraft
Affiliation:
University Department of Bacteriology, Glasgow Royal Infirmary, Castle Street, Glasgow G4 0SF
J. Taggart
Affiliation:
University Department of Bacteriology, Glasgow Royal Infirmary, Castle Street, Glasgow G4 0SF
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Restriction enzyme fingerprints were generated from purified plasmid DNA from 324 clinical isolates that belonged to 7 enterobacterial genera and 88 single plasmids in Escherichia coli K12 according to the following strategy.

Purified plasmid DNA was digested with PstI. The number of fragments detected in a 0·8 agarose gel was used to determine which 2 of 6 restriction enzymes including Pstl was most likely to provide a fingerprint comprising sufficient fragments to ensure specificity but sufficiently few to allow easy visual assessment and minimize coincidental matching. When PstI produced > 20 fragments, Eco RI and HindIII were used; when PstI generated < 6 fragments Bsp 1286 and AvaII were used and SmaI was employed when between 6 and 20 fragments were obtained from PstI digests. Using a minimum of 12 fragments from a combination of 2 enzymes as the criterion for characterizing a strain/plasmid, satisfactory 2-enzyme fingerprints were obtained from 87% of the strains and plasmids studied using PstI and no more than two additional enzymes per strain. Of the remaining 54 strains, 51 harboured only small plasmids (< 10 kb) and 3 produced satisfactory fingerprints when digested with a fourth enzyme.

Type
Research Article
Information
Journal of Hygiene , Volume 97 , Issue 2 , October 1986 , pp. 205 - 210
Copyright
Copyright © Cambridge University Press 1986

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 Baderoides fragilis. Journal of Medical Microbiology 20, 39–48.CrossRefGoogle ScholarPubMed
Callihan, D. R., Young, F. E. & Clark, V. L. (1983). Identification of three homology classes of small cryptic plasmids in intestinal Baderoides species. Plasmid 9, 17–30.CrossRefGoogle ScholarPubMed
Hawkey, P. M., Bennett, P. M. & Hawkey, C. A. (1984). Cryptic plasmids in hospital isolates of Providencia stuartii. Journal of Medical Microbiology 18, 277–284.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, 433–441.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, 95–105.CrossRefGoogle ScholarPubMed
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, 79–89.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., Christenson, 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, 1–6.CrossRefGoogle ScholarPubMed
Platt, D. J. (1983). Bacterial plasmids and their fingerprints. Disease Markers 1, 107–115.Google Scholar
Platt, D. J., Chesham, J. S. & Kristinsson, K. G. (1986). R-transfer in vivo: a prospective study. Journal of Medical Microbiology 21, 325–330.CrossRefGoogle ScholarPubMed
Platt, D. J. & Sommerville, J. S. (1981). Serratia species isolated from patients in a general hospital. Journal of Hospital Infection 2, 341–348.CrossRefGoogle ScholarPubMed
Richmond, M. H. & Petrocheilou, V. (1978). R-factor transfer in vivo in humans. In Microbiology (ed. Schlessinger, D.), pp. 273–276. Washington: ASM.Google Scholar
Rubens, C. E., Farrar, W. E. Jr, McGee, Z. A. & Schaffer, W. (1981). Evolution of a plasmid mediating resistance to multiple antimicrobial agents during a prolonged epidemic of nosocomial infections. Journal of Infections diseases 143, 170–181.CrossRefGoogle ScholarPubMed
Thompson, R., Hughes, S. G. & Broda, P. (1974). Plasmid identification using specific endonucleases. Molecular & General Genetics 133, 141–149.CrossRefGoogle ScholarPubMed