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Journal of Clinical Microbiology, November 2001, p. 3865-3870, Vol. 39, No. 11
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.11.3865-3870.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

PCR Analyses of tRNA Intergenic Spacer, 16S-23S Internal Transcribed Spacer, and Randomly Amplified Polymorphic DNA Reveal Inter- and Intraspecific Relationships of Enterobacter cloacae Strains

Maysa M. Clementino,1,* Ivano de Filippis,1 Carlos R. Nascimento,1 Regina Branquinho,1 Carmem L. Rocha,1 and Orlando B. Martins2

Department of Microbiology, National Institute of Quality Control in Health, FIOCRUZ,1 and Department of Medical Biochemistry, ICB/CCS, Federal University of Rio de Janeiro,2 Rio de Janeiro, Brazil

Received 21 February 2001/Returned for modification 25 April 2001/Accepted 13 August 2001

PCR analysis of tRNA intergenic spacer (tDNA-PCR) and of the 16S-23S internal transcribed spacer (ITS-PCR) and random amplified polymorphic DNA (RAPD) analysis were evaluated for their usefulness in characterization of Enterobacter cloacae strains isolated from both clinical origins and vaccine microbial contamination. tDNA-PCR presented specific and reproducible patterns for Enterobacter sakazakii ATCC 29004, Enterobacter aerogenes ATCC 13048, and Enterobacter cloacae ATCC 13047 and 23355 that presented the same profile for all 16 E. cloacae isolates, offering an alternative tool for species-level identification. ITS-PCR and RAPD analysis yielded completely different banding patterns for the 20 strains studied, except for E. cloacae strains isolated from different batches of vaccine that exhibited a unique pattern, suggesting contamination by the same strain. The combined use of tDNA-PCR and ITS-PCR in a one-step protocol allows accurate identification and typing of E. cloacae strains a few hours after the colony has been isolated.

* Corresponding author. Mailing address: Departamento de Microbiologia, Instituto Nacional de Controle da Qualidade em Saúde, FIOCRUZ, Avenida Brasil, 4365-Manguinhos, 21045-900 Rio de Janeiro-RJ, Brazil. Phone: 55-21-598-4290. Fax: 55-21-290-0915. E-mail: maysa{at}alpha.incqs.fiocruz.br.

Journal of Clinical Microbiology, November 2001, p. 3865-3870, Vol. 39, No. 11
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.11.3865-3870.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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Antimicrob. Agents Chemother. Clin. Microbiol. Rev.
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Copyright © 2001 by the American Society for Microbiology. All rights reserved.