Abstract
A total of 53 field and reference strains, including the type strains of the seven named species (nomenspecies) and belonging to the 18 described genomic species (DNA groups) of the genus Acinetobacter, were studied by amplified ribosomal DNA restriction analysis (ARDRA). Restriction analysis with the enzymes AluI, CfoI, MboI, RsaI, and MspI of the enzymatically amplified 16S rRNA genes allowed us to identify all species except the genomic species 4 (Acinetobacter haemolyticus) and 7 (A. johnsonii), 5 (A. junii) and 17, and 10 and 11, which clustered pairwise in three respective groups. Further analysis with the enzyme HaeIII, HinfI, NciI, ScrFI, or TaqI did not allow us to differentiate the species within these three clusters. However, use of a few additional simple phenotypic tests (hemolysis, growth at 37 degrees C, production of acid from glucose, and gelatin hydrolysis) can be used to differentiate between the species within these clusters. ARDRA proved to be a rapid and reliable method for the identification of most of the Acinetobacter genomic species, including the closely related DNA groups 1 (A. calcoaceticus), 2 (A. baumannii), 3, and 13. The results of this study suggest that ARDRA can be used for the identification of Acinetobacter species and as such may help to elucidate the ecology and clinical significance of the different species of this genus. Since ARDRA uses universal 16S rRNA gene primers, it is expected to be applicable to the identification of most bacterial species. Furthermore, ARDRA is less prone to contamination problems than PCR for detection, since the use of cultured organisms results in a large initial quantity of target DNA.
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- Bergogne-Bérézin E., Joly-Guillou M. L., Vieu J. F. Epidemiology of nosocomial infections due to Acinetobacter calcoaceticus. J Hosp Infect. 1987 Sep;10(2):105–113. doi: 10.1016/0195-6701(87)90135-6. [DOI] [PubMed] [Google Scholar]
- Bouvet P. J., Grimont P. A. Identification and biotyping of clinical isolates of Acinetobacter. Ann Inst Pasteur Microbiol. 1987 Sep-Oct;138(5):569–578. doi: 10.1016/0769-2609(87)90042-1. [DOI] [PubMed] [Google Scholar]
- Bouvet P. J., Jeanjean S. Delineation of new proteolytic genomic species in the genus Acinetobacter. Res Microbiol. 1989 May-Jun;140(4-5):291–299. doi: 10.1016/0923-2508(89)90021-1. [DOI] [PubMed] [Google Scholar]
- Deng S., Hiruki C., Robertson J. A., Stemke G. W. Detection by PCR and differentiation by restriction fragment length polymorphism of Acholeplasma, Spiroplasma, Mycoplasma, and Ureaplasma, based upon 16S rRNA genes. PCR Methods Appl. 1992 Feb;1(3):202–204. doi: 10.1101/gr.1.3.202. [DOI] [PubMed] [Google Scholar]
- Dijkshoorn L., Aucken H. M., Gerner-Smidt P., Kaufmann M. E., Ursing J., Pitt T. L. Correlation of typing methods for Acinetobacter isolates from hospital outbreaks. J Clin Microbiol. 1993 Mar;31(3):702–705. doi: 10.1128/jcm.31.3.702-705.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dijkshoorn L., Michel M. F., Degener J. E. Cell envelope protein profiles of Acinetobacter calcoaceticus strains isolated in hospitals. J Med Microbiol. 1987 Jun;23(4):313–319. doi: 10.1099/00222615-23-4-313. [DOI] [PubMed] [Google Scholar]
- Dijkshoorn L., van der Toorn J. Acinetobacter species: which do we mean? Clin Infect Dis. 1992 Oct;15(4):748–749. doi: 10.1093/clind/15.4.748. [DOI] [PubMed] [Google Scholar]
- Freney J., Bouvet P. J., Tixier C. Identification et détermination de la sensibilité aux antibiotiques de 31 souches cliniques d'Acinetobacter autres que A. baumannii. Ann Biol Clin (Paris) 1989;47(1):41–44. [PubMed] [Google Scholar]
- Gerner-Smidt P., Frederiksen W. Acinetobacter in Denmark: I. Taxonomy, antibiotic susceptibility, and pathogenicity of 112 clinical strains. APMIS. 1993 Nov;101(11):815–825. doi: 10.1111/j.1699-0463.1993.tb00186.x. [DOI] [PubMed] [Google Scholar]
- Gerner-Smidt P., Tjernberg I. Acinetobacter in Denmark: II. Molecular studies of the Acinetobacter calcoaceticus-Acinetobacter baumannii complex. APMIS. 1993 Nov;101(11):826–832. [PubMed] [Google Scholar]
- Gerner-Smidt P., Tjernberg I., Ursing J. Reliability of phenotypic tests for identification of Acinetobacter species. J Clin Microbiol. 1991 Feb;29(2):277–282. doi: 10.1128/jcm.29.2.277-282.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gurtler V., Wilson V. A., Mayall B. C. Classification of medically important clostridia using restriction endonuclease site differences of PCR-amplified 16S rDNA. J Gen Microbiol. 1991 Nov;137(11):2673–2679. doi: 10.1099/00221287-137-11-2673. [DOI] [PubMed] [Google Scholar]
- Henriksen S. D. Moraxella, Acinetobacter, and the Mimeae. Bacteriol Rev. 1973 Dec;37(4):522–561. doi: 10.1128/br.37.4.522-561.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jayarao B. M., Doré J. J., Jr, Oliver S. P. Restriction fragment length polymorphism analysis of 16S ribosomal DNA of Streptococcus and Enterococcus species of bovine origin. J Clin Microbiol. 1992 Sep;30(9):2235–2240. doi: 10.1128/jcm.30.9.2235-2240.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Joly-Guillou M. L., Bergogne-Berezin E., Vieu J. F. A study of the relationships between antibiotic resistance phenotypes, phage-typing and biotyping of 117 clinical isolates of Acinetobacter spp. J Hosp Infect. 1990 Jul;16(1):49–58. doi: 10.1016/0195-6701(90)90048-s. [DOI] [PubMed] [Google Scholar]
- Kämpfer P., Tjernberg I., Ursing J. Numerical classification and identification of Acinetobacter genomic species. J Appl Bacteriol. 1993 Sep;75(3):259–268. doi: 10.1111/j.1365-2672.1993.tb02775.x. [DOI] [PubMed] [Google Scholar]
- Rossau R., Duhamel M., Jannes G., Decourt J. L., Van Heuverswyn H. The development of specific rRNA-derived oligonucleotide probes for Haemophilus ducreyi, the causative agent of chancroid. J Gen Microbiol. 1991 Feb;137(2):277–285. doi: 10.1099/00221287-137-2-277. [DOI] [PubMed] [Google Scholar]
- Tjernberg I., Ursing J. Clinical strains of Acinetobacter classified by DNA-DNA hybridization. APMIS. 1989 Jul;97(7):595–605. doi: 10.1111/j.1699-0463.1989.tb00449.x. [DOI] [PubMed] [Google Scholar]
- Vaneechoutte M., De Beenhouwer H., Claeys G., Verschraegen G., De Rouck A., Paepe N., Elaichouni A., Portaels F. Identification of Mycobacterium species by using amplified ribosomal DNA restriction analysis. J Clin Microbiol. 1993 Aug;31(8):2061–2065. doi: 10.1128/jcm.31.8.2061-2065.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vaneechoutte M., Rossau R., De Vos P., Gillis M., Janssens D., Paepe N., De Rouck A., Fiers T., Claeys G., Kersters K. Rapid identification of bacteria of the Comamonadaceae with amplified ribosomal DNA-restriction analysis (ARDRA). FEMS Microbiol Lett. 1992 Jun 15;72(3):227–233. doi: 10.1111/j.1574-6968.1992.tb05102.x. [DOI] [PubMed] [Google Scholar]
- Weaver R. E., Actis L. A. Identification of Acinetobacter species. J Clin Microbiol. 1994 Jul;32(7):1833–1833. doi: 10.1128/jcm.32.7.1833-.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Welsh J., McClelland M. Genomic fingerprints produced by PCR with consensus tRNA gene primers. Nucleic Acids Res. 1991 Feb 25;19(4):861–866. doi: 10.1093/nar/19.4.861. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wiedmann-al-Ahmad M., Tichy H. V., Schön G. Characterization of Acinetobacter type strains and isolates obtained from wastewater treatment plants by PCR fingerprinting. Appl Environ Microbiol. 1994 Nov;60(11):4066–4071. doi: 10.1128/aem.60.11.4066-4071.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]