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Applied and Environmental Microbiology, November 2002, p. 5358-5366, Vol. 68, No. 11
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.11.5358-5366.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Differentiation of Closely Related Carnobacterium Food Isolates Based on 16S-23S Ribosomal DNA Intergenic Spacer Region Polymorphism

Petia Kabadjova, Xavier Dousset,* Virginie Le Cam, and Hervé Prevost

Laboratoire de Microbiologie Alimentaire et Industrielle, ENITIAA, F-44322 Nantes Cedex 3, France

Received 14 February 2002/ Accepted 5 August 2002

A novel strategy for identification of Carnobacterium food isolates based on restriction fragment length polymorphism (RFLP) of PCR-amplified 16S-23S ribosomal intergenic spacer regions (ISRs) was developed. PCR amplification from all Carnobacterium strains studied always yielded three ISR amplicons, which were designated the small ISR (S-ISR), the medium ISR (M-ISR), and the large ISR (L-ISR). The lengths of these ISRs varied from one species to another. Carnobacterium divergens NCDO 2763T and C. mobile DSM 4849T generated one major S-ISR band (ca. 400 bp) and minor M-ISR and L-ISR bands (ca. 500 and ca. 600 bp, respectively). The ISRs amplified from C. gallinarum NCFB 2766T and C. piscicola NCDO 2762T were larger (S-ISR, ca. 600 bp; M-ISR, ca. 700 bp; and L-ISR, ca. 800 bp). The L-ISR contained two tDNAs coding for tRNAIle and tRNAAla genes. The M-ISR included one tRNAAla gene, and the S-ISR did not contain a tDNA gene. The RFLP scheme devised involves estimation of variable PCR product sizes together with HinfI, TaqI, and HindIII restriction analysis. Forty-two isolates yielded four unique band patterns that correctly resolved these isolates into four Carnobacterium species. This method is very suitable for rapid, low-cost identification of a wide variety of Carnobacterium species without sequencing.

* Corresponding author. Mailing address: Laboratoire de Microbiologie Alimentaire et Industrielle, Département Sciences des Aliments, ENITIAA, rue de la Géraudière, BP 82225, 44322 Nantes Cedex 3, France. Phone: 33-2-51785525. Fax: 33-2-51785455. E-mail: dousset{at}enitiaa-nantes.fr.

Applied and Environmental Microbiology, November 2002, p. 5358-5366, Vol. 68, No. 11
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.11.5358-5366.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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