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Applied and Environmental Microbiology, May 2000, p. 2113-2116, Vol. 66, No. 5
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Multiplex PCR for the Detection of Lactobacillus pontis and Two Related Species in a Sourdough Fermentation

Martin R. A. Müller, Matthias A. Ehrmann,* and Rudi F. Vogel

Lehrstuhl für Technische Mikrobiologie, Technische Universität München, 85350 Freising, Germany

Received 20 September 1999/Accepted 11 February 2000

A specific multiplex PCR assay based on the amplification of parts of the 16S rRNA molecule was designed. Primers derived from variable regions of the 16S rRNA provided a means of easily differentiating the species Lactobacillus pontis and Lactobacillus panis. They could be clearly discriminated from the phylogenetically related species Lactobacillus vaginalis, Lactobacillus oris, and Lactobacillus reuteri and from other lactobacilli commonly known to be present in sourdough. Other strains isolated together with L. pontis from an industrial sourdough fermentation could be clearly separated from these species by comparative sequence analysis and construction of a specific PCR primer. For a fast identification a DNA isolation protocol based on the ultrasonic lysis of cells from single colonies was developed. To demonstrate the potential of such techniques for tracking these organisms in a laboratory-scale fermentation, we combined the specific PCR assay with direct DNA extraction from the organisms in the sourdough without previous cultivation.

* Corresponding author. Mailing address: Lehrstuhl für Technische Mikrobiologie, Weihenstephaner Steig 16, 85350 Freising, Germany. Phone: (49) 8161-715386. Fax: (49) 8161-713327. E-mail: M.Ehrmann{at}bl.tum.de.

Applied and Environmental Microbiology, May 2000, p. 2113-2116, Vol. 66, No. 5
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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