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Applied and Environmental Microbiology, January 2000, p. 297-303, Vol. 66, No. 1
0099-2240/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Detection and Identification of Gastrointestinal Lactobacillus Species by Using Denaturing Gradient Gel Electrophoresis and Species-Specific PCR Primers

J. Walter,1 G. W. Tannock,1,* A. Tilsala-Timisjarvi,2 S. Rodtong,3 D. M. Loach,1 K. Munro,1 and T. Alatossava4

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 27 July 1999/Accepted 28 October 1999

Denaturing gradient gel electrophoresis (DGGE) of DNA fragments obtained by PCR amplification of the V2-V3 region of the 16S rRNA gene was used to detect the presence of Lactobacillus species in the stomach contents of mice. Lactobacillus isolates cultured from human and porcine gastrointestinal samples were identified to the species level by using a combination of DGGE and species-specific PCR primers that targeted 16S-23S rRNA intergenic spacer region or 16S rRNA gene sequences. The identifications obtained by this approach were confirmed by sequencing the V2-V3 region of the 16S rRNA gene and by a BLAST search of the GenBank database.

* Corresponding author. Mailing address: Department of Microbiology, University of Otago, P.O. Box 56, Dunedin, New Zealand. Phone: 64-3-479-7734. Fax: 64-3-479-8540. E-mail: gerald.tannock{at}stonebow.otago.ac.nz.

Applied and Environmental Microbiology, January 2000, p. 297-303, Vol. 66, No. 1
0099-2240/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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