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Applied and Environmental Microbiology, June 2001, p. 2760-2765, Vol. 67, No. 6
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.6.2760-2765.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Specific Identification and Targeted Characterization of Bifidobacterium lactis from Different Environmental Isolates by a Combined Multiplex-PCR Approach

Marco Ventura, Roberto Reniero, and Ralf Zink*

Nestlé Research Center, Vers-Chez-Les-Blanc, 1000 Lausanne 26, Switzerland

Received 12 December 2000/Accepted 20 March 2001

The species Bifidobacterium lactis, with its main representative strain Bb12 (DSM 10140), is a yoghurt isolate used as a probiotic strain and is commercially applied in different types of yoghurts and infant formulas. In order to ensure the genetic identity and safety of this bacterial isolate, species- and strain-specific molecular tools for genetic fingerprinting must be available to identify isolated bifidobacteria or lactic acid bacteria from, e.g., various clinical environments of relevance in medical microbiology. Two opposing rRNA gene-targeted primers have been developed for specific detection of this microorganism by PCR. The specificity of this approach was evaluated and verified with DNA samples isolated from single and mixed cultures of bifidobacteria and lactobacilli (48 isolates, including the type strains of 29 Bifidobacterium and 9 Lactobacillus species). Furthermore, we performed a Multiplex-PCR using oligonucleotide primers targeting a specific region of the 16S rRNA gene for the genus Bifidobacterium and a conserved eubacterial 16S rDNA sequence. The specificity and sensitivity of this detection with a pure culture of B. lactis were, respectively, 100 bacteria/ml after 25 cycles of PCR and 1 to 10 bacteria/ml after a 50-cycle nested-PCR approach.

* Corresponding author. Mailing address: Nestlé Research Center, Route du Jourat 57, Vers-Chez-Les-Blanc, 1000 Lausanne 26, Switzerland. Phone: 41-21-785-8901. Fax: 41-21-785-8925. E-mail: ralf.zink{at}rdls.nestle.com.

Applied and Environmental Microbiology, June 2001, p. 2760-2765, Vol. 67, No. 6
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.6.2760-2765.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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