Movatterモバイル変換


[0]ホーム

URL:


Skip to main content

Advertisement

Springer Nature Link
Log in

DNA amplification fingerprinting (DAF) applied to the investigation ofAcinetobacter baumanii isolated from intensive care unit patients

  • Published:
Irish Journal of Medical Science Aims and scope Submit manuscript

Abstract

Acinetobacter are important emerging nosocomial pathogens. In this paper thirteenAcinetobacter baumanii from intensive care patients were isolated. These were initially typed using the API-20 NE biotyping system and antibiogram analysis. Results obtained using these methods failed to convincingly characterise the organisms. In this report a method to characterise theseAcinetobacter baumanii isolates is described, which utilises a modified polymerase chain reaction (PCR), capable of generating genomic fingerprints, known as DNA amplification fingerprinting (DAF). Purified chromosomal DNA of cultured clinical isolates ofAcinetobacter baumanii were subjected to DAF using the M13 universal sequencing primer. Polymorphic DNA bands produced, were visualised after agarose gel electrophoresis and ethiduim bromide staining. Results demonstrated that six of the thirteen clinical isolates represented one group and a second group of two isolates displayed identical fingerprint patterns. The remaining four organisms were all unique. This genotype based method is rapid, simple, reproducible and may have potential as a means of specifically typingAcinetobacter spp. allowing the route(s) of nosocomial transmission to be identified and to assess the efficiency of instituted infection control measures.

This is a preview of subscription content,log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+
from ¥17,985 /Month
  • Starting from 10 chapters or articles per month
  • Access and download chapters and articles from more than 300k books and 2,500 journals
  • Cancel anytime
View plans

Buy Now

Price includes VAT (Japan)

Instant access to the full article PDF.

Similar content being viewed by others

Explore related subjects

Discover the latest articles, books and news in related subjects, suggested using machine learning.

References

  1. Bouvet, P. J. M., Grimont, P. A. D. Taxonomy of the genusAcinetobacter with the recognition ofAcinetobacter baumanii sp. nov.,Acinetobacter haemolyticus sp. nov.,Acinetobacter johnsonii sp. nov., andAcinetobacter junii sp. nov. and amended discriptions ofAcinetobacter calcoaceticus andAcinetobacter lwoffi. Int. J. Syst. Bacteriol. 1986; 36: 228–240.

    Article CAS  Google Scholar 

  2. Kropec, A., Hubner, J., Daschner, F. D. Comparison of three typing methods in hospital outbreaks ofAcinetobacter calcoaceticus infection. J. Hosp. Infect. 1993; 23: 133–141.

    Article PubMed CAS  Google Scholar 

  3. Alexander, M., Rahman, M., Taylor, M., Noble, W. C. A. study of the value of electrophoretic and other techniques for typingAcinetobacter calcoaceticus. J. Hosp. Infect. 1988; 12: 273–287.

    Article PubMed CAS  Google Scholar 

  4. Mullis, K. B., Faloona, F. A. Specific synthesis of DNAin vitro via a polymerase-catalyzed chain reaction. Methods Enzymol. 1987; 155: 335–350.

    Article PubMed CAS  Google Scholar 

  5. Caetano-annollés, G., Bassam, B. J., Gresshoff, P. M. DNA amplification fingerprinting using very short arbitrary oligonucleotide primers. BioTechnology 1991; 9: 553–557.

    Article  Google Scholar 

  6. Palittapongarnpim, P., Chomyc, S., Fanning, A., Kunimoto, D. DNA fragment length polymorphism analysis ofMycobacteriunl tuberculosis isolates by arbitrarily primed polymerase chain reaction. J. Infect. Dis. 1993; 167: 975–978.

    PubMed CAS  Google Scholar 

  7. Welsh, J., McClelland, M. Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Res. 1990; 18: 7213–7218.

    Article PubMed CAS  Google Scholar 

  8. Williams, J. G. K., Kubelik, A. R., Livak, K. J., Rafalski, J. A., Tingey, S. V. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 1990; 18: 6531–6535.

    Article PubMed CAS  Google Scholar 

  9. Waterworth, P. M., Garrod, L. P., Lambert, H. P., O’Grady, F. Laboratory control in antibiotics and chemotherapy. 5th Edition, Edinburgh, Churchill Livingstone 1981; 464–504.

    Google Scholar 

  10. Bouvet, P. J. M., Grimont, P. A. D. Identification and biotyping of clinical isolates ofAcinetobacter. Ann. Inst. Pasteur. Microbiol. 1987; 138: 569–578.

    Article PubMed CAS  Google Scholar 

  11. Penner, G. A., Bush, A., Wise, K., Kim, W., Domier, L., Kasha, K., Laroche, A., Scoles, G., Molnar, S. J., Fedak, G. Reproducibility of random amplified polymorphic DNA (RAPD) analysis among laboratories. PCR Methods Applic. 1993; 2: 341–345.

    CAS  Google Scholar 

  12. Dijkshooln, L., van Dalen, R., van Ooyen, A., Bijl, D., Tjernberg, I., Michel, M. F., Horrevorts, A. M. Endemic acinetobacter in intensive care units:epidemiology and clinical impact. J. Clin. Pathol. 1993; 46: 533–536.

    Article  Google Scholar 

  13. Falkiner, F. R. Epidemiological typing: a user’s view. J. Hosp. infect. 1988; 11: 303–309.

    Article PubMed CAS  Google Scholar 

  14. Carlano, A. V., Lamerdin, J., Ashworth, L. K., Watkins, B., Branscomb, E., Slezak, T., Raff, M., DeJong, P. J., Kieth, D., McBride, L., Meister, S., Kronick, M. A. high-resolution, fluorescence-based, semi-automated method for DNA fingerprinting. Genomics 1989; 4: 129–136.

    Article  Google Scholar 

  15. Giusti, W. G., Adriano, T. Synthesis and characterization of 5’-fluorescent-dye-labeled oligonucleotides. PCR Methods Applic. 1993; 2: 223–227.

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Medical Sciences Section, Regional Technical College, Rossa Avenue, Bishopstown, Cork, Ireland

    C. Sheehan, L. Boissel & S. Fanning

  2. Department of Medical Microbiology, Cork University Hospital Cork, Ireland

    M. Lynch & B. Cryan

  3. Department of Molecular and Human Genetics, Baylor College of Medicine, 77030, Houston, Texas

    S. Fanning

Authors
  1. C. Sheehan
  2. L. Boissel
  3. M. Lynch
  4. B. Cryan
  5. S. Fanning

Corresponding author

Correspondence toS. Fanning.

Rights and permissions

About this article

Access this article

Subscribe and save

Springer+
from ¥17,985 /Month
  • Starting from 10 chapters or articles per month
  • Access and download chapters and articles from more than 300k books and 2,500 journals
  • Cancel anytime
View plans

Buy Now

Price includes VAT (Japan)

Instant access to the full article PDF.

Advertisement


[8]ページ先頭

©2009-2026 Movatter.jp