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Applied and Environmental Microbiology, September 1999, p. 4264-4267, Vol. 65, No. 9
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Identification of Lactobacillus Isolates from the Gastrointestinal Tract, Silage, and Yoghurt by 16S-23S rRNA Gene Intergenic Spacer Region Sequence Comparisons

G. W. Tannock,1,* A. Tilsala-Timisjarvi,2 S. Rodtong,3 J. Ng,1 K. Munro,1 and T. Alatossava2,4

Department of Microbiology, University of Otago, Dunedin, New Zealand1; Department of Biology, University of Oulu, Oulu,2 and Biotechnology Laboratory, REDEC of Kajaani, University of Oulu, Sotkamo,4 Finland; and School of Microbiology, Suranaree University of Technology, Nakhon Ratchasima, Thailand3

Received 16 February 1999/Accepted 1 July 1999

Lactobacillus isolates were identified by PCR amplification and sequencing of the region between the 16S and 23S rRNA genes (spacer region). The sequences obtained from the isolates were compared to those of reference strains held in GenBank. A similarity of 97.5% or greater was considered to provide identification. To check the reliability of the method, the V2-V3 region of the 16S rRNA gene was amplified and sequenced in the case of isolates whose spacer region sequences were less than 99% similar to that of a reference strain. Confirmation of identity was obtained in all instances. Spacer region sequencing provided rapid and accurate identification of Lactobacillus isolates obtained from gastrointestinal, yoghurt, and silage samples. It had an advantage over 16S V2-V3 sequence comparisons because it distinguished between isolates of Lactobacillus casei and Lactobacillus rhamnosus.

* Corresponding author. Mailing address: Department of Microbiology, University of Otago, P.O. Box 56, Dunedin, New Zealand. Phone: 64-3-479-7713. Fax: 64-3-479-8540. E-mail: gerald.tannock{at}stonebow.otago.ac.nz.

Applied and Environmental Microbiology, September 1999, p. 4264-4267, Vol. 65, No. 9
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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