Antonie van Leeuwenhoek

, Volume 88, Issue 2, pp 173–179 | Cite as

BOX-PCR is an Adequate Tool for Typing Aeromonas spp.

  • Marta Tacão
  • Artur Alves
  • Maria José Saavedra
  • António Correia
Short Communication


PCR-based methods of fingerprinting take advantage of the presence of repetitive sequences that are interspersed throughout the genome of diverse bacterial species. They include the repetitive extragenic palindromic (REP) sequence, the enterobacterial repetitive intergenic consensus sequence (ERIC) and the 154-bp BOX element. The combination of the three methods is used for fine discrimination of strains and is designated as rep-polymerase chain reaction (PCR). REP-PCR and ERIC-PCR have been shown to be useful for typing Aeromonas strains. To our knowledge, rep-PCR fingerprinting method using the BOXA1R primer has never been tested on aeromonads. In this study, the BOX-PCR fingerprinting technique was evaluated for the discrimination of strains of some Aeromonas species. All strains were typeable and the majority showed unique banding patterns. Four strains from culture collections were used to investigate the reproducibility of the method. According to our results, BOX-PCR fingerprinting is applicable for typing of Aeromonas strains and can be considered as a useful complementary tool for epidemiological studies of members of this genus.


Aeromonas spp. BOX-PCR Molecular typing 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Altschul, S.F., Madden, T.L., Schäffer, A.A., Zhang, J., Zhang, Z., Miller, W., Lipman, D.J. 1997Gapped BLAST and PSI-BLAST: a new generation of protein database search programsNucleic Acids Res.2533893402CrossRefPubMedGoogle Scholar
  2. Altwegg, M. 1996Subtyping methods for Aeromonas speciesAustin, B.Altwegg, M.Gosling, P.J.Joseph, S.W. eds. The Genus AeromonasJohn Wiley and sonsChichesterUnited Kingdom109126Google Scholar
  3. Altwegg, M., Reeves, M.W., Altwegg-Bissig, R., Brenner, D.J. 1991Multilocus enzyme analysis of the genus Aeromonasits use for species identificationZentralbl. Bakteriol.2752845PubMedGoogle Scholar
  4. Bonadonna, L., Briancesco, R., Filetici, E., Manuppella, A., Pourshaban, M., Semproni, M. 2002Genomic heterogeneity of environmental and clinical aeromonadsMicrobiologica252129PubMedGoogle Scholar
  5. Borrell, N., Acinas, S.G., Figueras, M.J., Martinez-Murcia, A. 1997Identification of Aeromonas clinical isolates by restriction fragment length polymorphism of PCR-amplified 16S rRNA genesJ. Clin. Microbiol.3516711674PubMedGoogle Scholar
  6. Brandi, G., Sisti, M., Schiavano, G.F., Salvaggio, L., Albano, A. 1996Survival of Aeromonas hydrophilaAeromonas caviaeAeromonas sobria in soilJ. Appl. Bacteriol.81439444Google Scholar
  7. DavinRegli, A., Bollet, C., Chamorey, E., Distria, V.C., Cremieux, A. 1998A cluster of cases of infections due to Aeromonas hydrophila revealed by combined RAPD and ERIC-PCRJ. Med. Microbiol.47499504PubMedGoogle Scholar
  8. Figueras, M.J., Guarro, J., Martínez-Murcia, A. 2000Use of restriction fragment length polymorphism of the PCR-amplified 16S rRNA gene for the identification of Aeromonas sppJ. Clin. Microbiol.3820232025PubMedGoogle Scholar
  9. Hänninen, M.L., Hirvelä-Kosky, V. 1997Pulsed-field electrophoresis in the study of mesophilic and psychrophilic Aeromonas sppJ Appl. Microbiol.83493498CrossRefGoogle Scholar
  10. Harf-Monteil, C., LeFlèche, A., Riegel, P., Prévost, G., Bermond, D., Grimont, P.A.D., Monteil, H. 2004Aeromonas simiae sp. nov., isolated from monkey faecesInt. J. Syst. Evol. Microbiol.54481485CrossRefPubMedGoogle Scholar
  11. Holmes, P., Niccols, L.M., Sartory, D.P. 1996The ecology of mesophilic Aeromonas in the aquatic environmentAustin, B.Altwegg, M.Gosling, P.J.Joseph, S. eds. The Genus AeromonasJohn Wiley and SonsChichesterEngland127150Google Scholar
  12. Janda, J.M., Abbott, S.L. 1998Evolving concepts regarding the genus Aeromonas: an expanding panorama of species, disease presentations, and unanswered questionsClin. Infect. Dis.27332344PubMedGoogle Scholar
  13. Martínez-Murcia, A.J., Borrell, N., Figueras, M.J. 2000Typing of clinical and environmental Aeromonas veronii strains based on the 16S–23S rDNA spacersFEMS Immunol. Med. Microbiol.28225232CrossRefPubMedGoogle Scholar
  14. Miñana-Galbis, D., Farfán, M., Fusté, M.C., Lorén, J.G. 2004Aeromonas molluscorum sp. nov. isolated from bivalve molluscsInt. J. Syst. Evol. Microbiol.5420732078CrossRefPubMedGoogle Scholar
  15. Moyer, N.P., Martinetti, G., Lüthy-Hottenstein, J., Altwegg, M. 1992Value of rRNA gene restriction patterns of Aeromonas spp. for epidemiological investigationsCurr. Microbiol.241521CrossRefGoogle Scholar
  16. Nei, M., Li, W.H. 1979Mathematical model for studying genetic variation in terms of restriction endonucleasesProc. Natl. Acad. Sci. USA7652695273PubMedGoogle Scholar
  17. Oakey, H.J., Gibson, L.F., George, A.M. 1998Co-migration of RAPD-PCR amplicons from Aeromonas hydrophilaFEMS Microbiol. Lett.1643538CrossRefPubMedGoogle Scholar
  18. Pidiyar, V., Kaznowski, A., Badri Narayan, N., Patole, M., Shouche, Y.S. 2002Aeromonas culicicola sp. nov., from the midgut of Culex quinquefasciatusInt. J. Syst. Evol. Microbiol.5217231728CrossRefPubMedGoogle Scholar
  19. Popoff, M. 1984Genus III. Aeromonas Kluyver and Van Niel 1936, 398ALKrieg, N.R.Holt, J.G. eds. Bergey’s Manual of Systematic Bacteriology, Vol. 1Williams and Wilkins Co.BaltimoreUSA545548Google Scholar
  20. Rademaker, J.L.W., de Bruijn, F.J. 1997Characterization and classification of microbes by rep-PCR genomic fingerprinting and computer assisted pattern analysisCaetano-Anollés, G.Gresshoff, P.M. eds. DNA Markers: Protocols, Applications and OverviewsJohn Wiley and Sons Inc.New York151171Google Scholar
  21. Rohlf, F.J. 1993NTSYS-pc. Numerical Taxonomy and Multivariate Analysis SystemExeter SoftwareNew York, USAGoogle Scholar
  22. Soler, L., Figueras, M.J., Chacón, M.R., Guarro, J., Martinez-Murcia, A.J. 2003Comparison of three molecular methods for typing Aeromonas popoffii isolatesAntonie van Leeuwenhoek795359Google Scholar
  23. Szczuka, E., Kaznowski, A. 2004Typing of clinical and environmental Aeromonas sp. strains by random amplified polymorphic DNA PCR, repetitive extragenic palindromic PCR, and enterobacterial repetitive intergenic consensus sequence PCRJ. Clin. Microbiol.42220228CrossRefPubMedGoogle Scholar
  24. Talon, D., Dupont, M.J., Thouverez, M., Michel-Briand, Y. 1996Pulsed-field gel electrophoresis as an epidemiological tool for clonal identification of Aeromonas hydrophilaJ. Appl. Bacteriol.80277282PubMedGoogle Scholar
  25. Versalovic, J., Schneider, M., de Bruijn, F.J., Lupski, J.R. 1994Genomic fingerprinting of bacteria using the repetitive sequence-based polymerase chain reactionMethods Mol. Cell. Biol.52540Google Scholar
  26. Villari, P., Crispino, M., Montuori, P., Bocciai, S. 2003Molecular typing of Aeromonas isolates in natural mineral watersAppl. Environ. Microbiol.69697701CrossRefPubMedGoogle Scholar
  27. Weisburg, W.G., Barns, S.M., Pelletier, D.A., Lane, D.J. 199116S ribosomal DNA amplification for phylogenetic studyJ. Bacteriol.173697703PubMedGoogle Scholar

Copyright information

© Springer 2005

Authors and Affiliations

  • Marta Tacão
    • 1
  • Artur Alves
    • 1
  • Maria José Saavedra
    • 2
  • António Correia
    • 1
  1. 1.Department of Biology, Center for Cell BiologyUniversity of AveiroAveiroPortugal
  2. 2.CECAV – Department of Veterinary ScienceUniversity of Trás-os-Montes e Alto DouroVila RealPortugal

Personalised recommendations