Typing Acinetobacter Strains: Applications and Methods

  • Lenie Dijkshoorn
Part of the Infectious Agents and Pathogenesis book series (IAPA)


The emergence of acinetobacters, with Acinetobacter baumannii in particular, as nosocomial pathogens has raised questions about their diversity, and the sources and mode of spread of particular strains. These questions cannot be answered without the availability of methods for strain typing. Over the past decades, there has been an explosive development of typing methods and many of these have been instrumental in the study of acinetobacters, not only for outbreak analysis but also for other purposes. The present paper provides an overview of applications of methods in the study of these organisms.

Applications of Methods for Typing Acinetobacter Strains

The term ‘typing’ refers to the activity to differentiate bacterial strains below species level. Typing of acinetobacters can be done for a variety of reasons (Table 1). For example, many studies have been performed to evaluate new typing methods, to identify epidemic strains, to study the geographic spread of particular...


Genomic Species Epidemic Strain PFGE Typing Enterobacterial Repetitive Intergenic Consensus Repetitive Extragenic Palindromic 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



Tanny van der Reijden and Beppie van Strijen are thanked for the preparation of pictures and Alexandr Nemec for critical reading of the text.


  1. Allardet-Servent A., Bouziges N., Carles-Nurit M.J., Bourg G., Gouby A., and Ramuz M. 1989. Use of low-frequency-cleavage restriction endonucleases for DNA analysis in epidemiological investigations of nosocomial bacterial infections. J. Clin. Microbiol. 27:2057–2061.PubMedGoogle Scholar
  2. Ayats J., Corbella X., Ardanuy C., Dominguez M.A., Ricart A., Ariza J., Martin R., and Linares J. 1997. Epidemiological significance of cutaneous, pharyngeal, and digestive tract colonization by multi-resistant Acinetobacter baumannii in ICU patients. J. Hosp. Infect. 37:287–295.PubMedCrossRefGoogle Scholar
  3. Bartual S.G., Seifert H., Hippler C., Luzon M.A., Wisplinghoff H., and Rodriguez-Valera F. 2005. Development of a multi-locus sequence typing scheme for characterization of clinical isolates of Acinetobacter baumannii. J. Clin. Microbiol. 43:4382–4390.PubMedCrossRefGoogle Scholar
  4. Bernards A.T., de Beaufort A.J., Dijkshoorn L., and van Boven C.P. 1997. Outbreak of septicaemia in neonates caused by Acinetobacter junii investigated by amplified ribosomal DNA restriction analysis (ARDRA) and four typing methods. J. Hosp. Infect. 35:129–140.PubMedCrossRefGoogle Scholar
  5. Bouvet P.J., and Grimont P.A. 1987. Identification and biotyping of clinical isolates of Acinetobacter. Ann. Inst. Pasteur Microbiol. 138:569–578.PubMedCrossRefGoogle Scholar
  6. Bouvet P.J., Jeanjean S., Vieu J.F., and Dijkshoorn L. 1990. Species, biotype, and bacteriophage type determinations compared with cell envelope protein profiles for typing Acinetobacter strains. J. Clin. Microbiol. 28:170–176.PubMedGoogle Scholar
  7. Brisse S., Milatovic D., Fluit A.C., Kusters K., Toelstra A., Verhoef J., and Schmitz F.J. 2000. Molecular surveillance of European quinolone-resistant clinical isolates of Pseudomonas aeruginosa and Acinetobacter spp. using automated ribotyping. J. Clin. Microbiol. 38:3636–3645.PubMedGoogle Scholar
  8. Da Silva G., Dijkshoorn L., Van Der R.T., van S.B., and Duarte A. 2007. Identification of widespread, closely related Acinetobacter baumannii isolates in Portugal as a subgroup of European clone II. Clin. Microbiol. Infect. 13:190–195.PubMedGoogle Scholar
  9. Dijkshoorn L., Aucken H., Gerner-Smidt P., Janssen P., Kaufmann M.E., Garaizar J., Ursing J., and Pitt T.L. 1996. Comparison of outbreak and non-outbreak Acinetobacter baumannii strains by genotypic and phenotypic methods. J. Clin. Microbiol. 34:1519–1525.PubMedGoogle Scholar
  10. Dijkshoorn L., Aucken H.M., Gerner-Smidt P., Kaufmann M.E., Ursing J., and Pitt T.L. 1993. Correlation of typing methods for Acinetobacter isolates from hospital outbreaks. J. Clin. Microbiol. 31:702–705.PubMedGoogle Scholar
  11. Dijkshoorn L., Tjernberg I., Pot B., Michel M.F., Ursing J., and Kersters K. 1990. Numerical analysis of cell envelope protein profiles of Acinetobacter strains classified by DNA-DNA hybridization. Syst. Appl. Microbiol. 13:338–334.CrossRefGoogle Scholar
  12. Dijkshoorn L., Van Vianen W., Degener J.E. and Michel M.F. 1987. Typing of Acinetobacter calcoaceticus strains isolated from hospital patients by cell envelope protein profiles. Epidemiol. Infect. 99:659–667.PubMedCrossRefGoogle Scholar
  13. Ecker J.A., Massire C., Hall T.A., Ranken R., Pennella T.T., Agasino I.C., Blyn L.B., Hofstadler S.A., Endy T.P., Scott P.T., Lindler L., Hamilton T., Gaddy C., Snow K., Pe M., Fishbain J., Craft D., Deye G., Riddell S., Milstrey E., Petruccelli B., Brisse S., Harpin V., Schink A., Ecker D.J., Sampath R., and Eshoo M.W. 2006. Identification of Acinetobacter species and genotyping of Acinetobacter baumannii by multilocus PCR and mass spectrometry. J. Clin. Microbiol. 44:2921–2932.PubMedCrossRefGoogle Scholar
  14. Ellington M.J., Kistler J., Livermore D.M., and Woodford N. 2007. Multiplex PCR for rapid detection of genes encoding acquired metallo-beta-lactamases. J. Antimicrob. Chemother. 59:321–322.PubMedCrossRefGoogle Scholar
  15. Enright M.C., and Spratt B.G. 1999. Multilocus sequence typing. Trends Microbiol. 7:482–487.PubMedCrossRefGoogle Scholar
  16. Freeman R., Sisson P.R., Noble W.C., and Lightfoot N.F. 1997. An apparent outbreak of infection with Acinetobacter calcoaceticus reconsidered after investigation by pyrolysis mass spectrometry. Zentralbl. Bakteriol. 285:234–244.PubMedCrossRefGoogle Scholar
  17. Gerner-Smidt P. 1989. Frequency of plasmids in strains of Acinetobacter calcoaceticus. J. Hosp. Infect. 14:23–28.PubMedCrossRefGoogle Scholar
  18. Gerner-Smidt P. 1992. Ribotyping of the Acinetobacter calcoaceticusAcinetobacter baumannii complex. J. Clin. Microbiol. 30:2680–2685.PubMedGoogle Scholar
  19. Gerner-Smidt P., and Tjernberg I. 1993. Acinetobacter in Denmark: II. Molecular studies of the Acinetobacter calcoaceticusAcinetobacter baumannii complex. APMIS 101:826–832.PubMedCrossRefGoogle Scholar
  20. Giammanco A., Vieu J.F., Bouvet P.J., Sarzana A., and Sinatra A. 1989. A comparative assay of epidemiological markers for Acinetobacter strains isolated in a hospital. Zentralbl. Bakteriol. 272:231–241.PubMedCrossRefGoogle Scholar
  21. Goncalves C.R., Vaz T.M., Araujo E., Boni R., Leite D., and Irino K. 2000. Biotyping, serotyping and ribotyping as epidemiological tools in the evaluation of Acinetobacter baumannii dissemination in hospital units, Sorocaba, Sao Paulo, Brazil. Rev. Inst. Med. Trop. Sao Paulo 42:277–282.PubMedCrossRefGoogle Scholar
  22. Gouby A., Carles-Nurit M.J., Bouziges N., Bourg G., Mesnard R., and Bouvet P.J. 1992. Use of pulsed-field gel electrophoresis for investigation of hospital outbreaks of Acinetobacter baumannii. J. Clin. Microbiol. 30:1588–1591.PubMedGoogle Scholar
  23. Grundmann H.J., Towner K.J., Dijkshoorn L., Gerner-Smidt P., Maher M., Seifert H., and Vaneechoutte M. 1997. Multicenter study using standardized protocols and reagents for evaluation of reproducibility of PCR-based fingerprinting of Acinetobacter spp. J. Clin. Microbiol. 35:3071–3077.PubMedGoogle Scholar
  24. Huys G., Cnockaert M., Nemec A., Dijkshoorn L., Brisse S., Vaneechoutte M., and Swings J. 2005a. Repetitive-DNA-element PCR fingerprinting and antibiotic resistance of pan-European multi-resistant Acinetobacter baumannii clone III strains. J. Med. Microbiol. 54:851–856.CrossRefGoogle Scholar
  25. Huys G., Cnockaert M., Nemec A., and Swings J. 2005b. Sequence-based typing of adeB as a potential tool to identify intraspecific groups among clinical strains of multidrug-resistant Acinetobacter baumannii. J. Clin. Microbiol. 43:5327–5331.CrossRefGoogle Scholar
  26. Janssen P., Coopman R., Huys G., Swings J., Bleeker M., Vos P., Zabeau M., and Kersters K. 1996. Evaluation of the DNA fingerprinting method AFLP as an new tool in bacterial taxonomy. Microbiology 142:1881–1893.PubMedCrossRefGoogle Scholar
  27. Janssen P., and Dijkshoorn L. 1996. High resolution DNA fingerprinting of Acinetobacter outbreak strains. FEMS Microbiol. Lett. 142:191–194.PubMedCrossRefGoogle Scholar
  28. Janssen P., Maquelin K., Coopman R., Tjernberg I., Bouvet P., Kersters K., and Dijkshoorn L. 1997. Discrimination of Acinetobacter genomic species by AFLP fingerprinting. Int. J. Syst. Bacteriol. 47:1179–1187.PubMedCrossRefGoogle Scholar
  29. Joly-Guillou M.L., Bergogne-Berezin E., and Vieu J.F. 1990. A study of the relationships between antibiotic resistance phenotypes, phage-typing and biotyping of 117 clinical isolates of Acinetobacter spp. J. Hosp. Infect. 16:49–58.PubMedCrossRefGoogle Scholar
  30. Koeleman J.G., Stoof J., Biesmans D.J., Savelkoul P.H., and Vandenbroucke-Grauls C.M. 1998. Comparison of amplified ribosomal DNA restriction analysis, random amplified polymorphic DNA analysis, and amplified fragment length polymorphism fingerprinting for identification of Acinetobacter genomic species and typing of Acinetobacter baumannii. J. Clin. Microbiol. 36:2522–2529.PubMedGoogle Scholar
  31. Liu P.Y., and Wu W.L. 1997. Use of different PCR-based DNA fingerprinting techniques and pulsed-field gel electrophoresis to investigate the epidemiology of Acinetobacter calcoaceticusAcinetobacter baumannii complex. Diagn. Microbiol. Infect. Dis. 29:19–28.PubMedCrossRefGoogle Scholar
  32. Maquelin K., Dijkshoorn L., Van Der Reijden T.J., and Puppels G.J. 2005. Rapid epidemiological analysis of Acinetobacter strains by Raman spectroscopy. J. Microbiol. Methods.Google Scholar
  33. Marchaim D., Navon-Venezia S., Schwartz D., Tarabeia J., Fefer I., Schwaber M.J., and Carmeli Y. 2007. Surveillance cultures and duration of carriage of multi-drug-resistant Acinetobacter baumannii. J. Clin. Microbiol 45:1551–1555.Google Scholar
  34. Nemec A., De Baere T., Tjernberg I., Vaneechoutte M., van der Reijden T.J., and Dijkshoorn L. 2001. Acinetobacter ursingii sp. nov. and Acinetobacter schindleri sp. nov., isolated from human clinical specimens. Int. J. Syst. Evol. Microbiol. 51:1891–1899.PubMedCrossRefGoogle Scholar
  35. Nemec A., Dijkshoorn L., and van der Reijden T.J. 2004a. Long-term predominance of two pan-European clones among multi-resistant Acinetobacter baumannii strains in the Czech Republic. J. Med. Microbiol. 53:147–153.CrossRefGoogle Scholar
  36. Nemec A., Dolzani L., Brisse S., van den Broek, P., and Dijkshoorn L. 2004b. Diversity of aminoglycoside-resistance genes and their association with class 1 integrons among strains of pan-European Acinetobacter baumannii clones. J. Med. Microbiol. 53:1233–1240.CrossRefGoogle Scholar
  37. Nemec A., Janda L., Melter O., and Dijkshoorn L. 1999. Genotypic and phenotypic similarity of multiresistant Acinetobacter baumannii isolates in the Czech Republic. J. Med. Microbiol. 48:287–296.PubMedCrossRefGoogle Scholar
  38. Nemec A., Maixnerova M., van der Reijden T.J., van den Broek P.J., and Dijkshoorn L. 2007. Relationship between the AdeABC efflux system gene content, netilmicin susceptibility and multidrug resistance in a genotypically diverse collection of Acinetobacter baumannii strains. J. Antimicrob. Chemother. Jun 26; [Epub ahead of print].Google Scholar
  39. Olive D.M., and Bean P. 1999. Principles and applications of methods for DNA-based typing of microbial organisms. J. Clin. Microbiol. 37:1661–1669.PubMedGoogle Scholar
  40. Orskov F., and Orskov I. 1983. From the national institutes of health. Summary of a workshop on the clone concept in the epidemiology, taxonomy, and evolution of the enterobacteriaceae and other bacteria. J. Infect. Dis. 148:346–357.PubMedCrossRefGoogle Scholar
  41. Pantophlet R., Nemec A., Brade L., Brade H., and Dijkshoorn L. 2001. O-antigen diversity among Acinetobacter baumannii strains from the Czech Republic and Northwestern Europe, as determined by lipopolysaccharide-specific monoclonal antibodies. J. Clin. Microbiol. 39:2576–2580.PubMedCrossRefGoogle Scholar
  42. Seifert H., Dolzani L., Bressan R., van der Reijden T., van Strijen B., Stefanik D., Heersma H., and Dijkshoorn L. 2005. Standardization and interlaboratory reproducibility assessment of pulsed-field gel electrophoresis-generated fingerprints of Acinetobacter baumannii. J. Clin. Microbiol. 43:4328–4335.PubMedCrossRefGoogle Scholar
  43. Seifert H., and Gerner-Smidt P. 1995. Comparison of ribotyping and pulsed-field gel electrophoresis for molecular typing of Acinetobacter isolates. J. Clin. Microbiol. 33:1402–1407.PubMedGoogle Scholar
  44. Seifert H., Schulze A., Baginski R., and Pulverer G. 1994a. Comparison of four different methods for epidemiologic typing of Acinetobacter baumannii. J. Clin. Microbiol. 32:1816–1819.Google Scholar
  45. Seifert H., Schulze A., Baginski R., and Pulverer G. 1994b. Plasmid DNA fingerprinting of Acinetobacter species other than Acinetobacter baumannii. J. Clin. Microbiol. 32:82–86.Google Scholar
  46. Singh A., Goering R.V., Simjee S., Foley S.L., and Zervos M.J. 2006. Application of molecular techniques to the study of hospital infection. Clin. Microbiol. Rev. 19:512–530.PubMedCrossRefGoogle Scholar
  47. Towner K.J., and Chopade B.A. 1987. Biotyping of Acinetobacter calcoaceticus using the API 2ONE system. J. Hosp. Infect. 10:145–151.PubMedCrossRefGoogle Scholar
  48. Traub W.H. 2000. Examination of polyclonal rabbit immune sera against serovars of Acinetobacter baumannii and genospecies 3 for cross-reactions with reference strains of other named/unnamed genospecies of Acinetobacter. Zentralbl. Bakteriol. 289:787–795.PubMedCrossRefGoogle Scholar
  49. Turton J.F., Gabriel S.N., Valderrey C., Kaufmann M.E., and Pitt T.L. 2007. Use of sequence-based typing and multiplex PCR to identify clonal lineages of outbreak strains of Acinetobacter baumannii. Clin. Microbiol. Infect. 13:807–815.PubMedCrossRefGoogle Scholar
  50. Turton J.F., Kaufmann M.E., Glover J., Coelho J.M., Warner M., Pike R., and Pitt T.L. 2005. Detection and typing of integrons in epidemic strains of Acinetobacter baumannii found in the United Kingdom. J. Clin. Microbiol. 43:3074–3082.PubMedCrossRefGoogle Scholar
  51. Turton J.F., Kaufmann M.E., Warner M., Coelho J., Dijkshoorn L., van der Reijden T., and Pitt T.L. 2004. A prevalent, multiresistant clone of Acinetobacter baumannii in Southeast England. J. Hosp. Infect. 58:170–179.PubMedCrossRefGoogle Scholar
  52. Van Belkum, A., Tassio P.T., Dijkshoorn L., Haeggman S, Cookson B, Fry N.K., Fussing V., Green J., Gerner-Smidt P., Brisse S., and Struelens M. 2007. European Society for Clinical Microbiology and Infectious Diseases (ESCMID) Study Group on Epidemiological Markers (ESGEM). CMI, Clin. Microbiol. Infect. 13(suppl, 3):1–46.Google Scholar
  53. Van Dessel H., Dijkshoorn L., Van Der Reijden T., Bakker N., Paauw A., Van Den B.P., Verhoef J., and Brisse S. 2004. Identification of a new geographically widespread multiresistant Acinetobacter baumannii clone from European hospitals. Res. Microbiol. 155:105–112.PubMedCrossRefGoogle Scholar
  54. Valenzuela J.K., Thomas L., Partridge S.R., van der Reijden T., Dijkshoorn L., and Iredell J. 2007. Horizontal gene transfer in a polyclonal outbreak of carbapenem-resistant Acinetobacter baumannii. J. Clin. Microbiol. 45:453–460. Epub 2006; Nov 15.PubMedCrossRefGoogle Scholar
  55. Vaneechoutte M., Elaichouni A., Maquelin K., Claeys G., Van Liedekerke A., Louagie H., Verschraegen G., and Dijkshoorn L. 1995. Comparison of arbitrarily primed polymerase chain reaction and cell envelope protein electrophoresis for analysis of Acinetobacter baumannii and A. junii outbreaks. Res. Microbiol. 146:457–465.PubMedCrossRefGoogle Scholar
  56. Vila J., Marcos M.A., and Jimenez de Anta M.T. 1996. A comparative study of different PCR-based DNA fingerprinting techniques for typing of the Acinetobacter calcoaceticusA. baumannii complex. J. Med. Microbiol. 44:482–489.PubMedCrossRefGoogle Scholar
  57. Weernink A., Severin W.P., Tjernberg I., and Dijkshoorn L. 1995. Pillows, an unexpected source of Acinetobacter. J. Hosp. Infect. 29:189–199.PubMedCrossRefGoogle Scholar
  58. Wroblewska M.M., Dijkshoorn L., Marchel H., van den B.M., Swoboda-Kopec E., van den Broek P.J., and Luczak M. 2004. Outbreak of nosocomial meningitis caused by Acinetobacter baumannii in neurosurgical patients. J. Hosp. Infect. 57:300–307.PubMedCrossRefGoogle Scholar
  59. Wu T.L., Ma L., Chang J.C., Su L.H., Chu C., Leu H.S., and Siu L.K. 2004. Variable resistance patterns of integron-associated multi-drug-resistant Acinetobacter baumannii isolates in a surgical intensive care unit. Microb. Drug Resist. 10:292–299.PubMedCrossRefGoogle Scholar

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© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  1. 1.Department of Infectious DiseasesLeiden University Medical CenterAlbinusdreef 2, Postbus 9600The Netherlands

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