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Applied and Environmental Microbiology, October 1999, p. 4351-4356, Vol. 65, No. 10
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Use of PCR-Based Methods for Rapid Differentiation of Lactobacillus delbrueckii subsp. bulgaricus and L. delbrueckii subsp. lactis

Sandra Torriani,1,2 Giacomo Zapparoli,1 and Franco Dellaglio1,*

Dipartimento Scientifico e Tecnologico, Facoltà di Scienze MM.FF.NN., Università degli Studi di Verona, 37134 Verona,1 and Istituto di Industrie Agrarie, Università degli Studi di Catania, 95124 Catania,2 Italy

Received 28 September 1998/Accepted 30 June 1999

Two PCR-based methods, specific PCR and randomly amplified polymorphic DNA PCR (RAPD-PCR), were used for rapid and reliable differentiation of Lactobacillus delbrueckii subsp. bulgaricus and L. delbrueckii subsp. lactis. PCR with a single combination of primers which targeted the proline iminopeptidase (pepIP) gene of L. delbrueckii subsp. bulgaricus allowed amplification of genomic fragments specific for the two subspecies when either DNA from a single colony or cells extracted from dairy products were used. A numerical analysis of the RAPD-PCR patterns obtained with primer M13 gave results that were consistent with the results of specific PCR for all strains except L. delbrueckii subsp. delbrueckii LMG 6412T, which clustered with L. delbrueckii subsp. lactis strains. In addition, RAPD-PCR performed with primer 1254 provided highly polymorphic profiles and thus was superior for distinguishing individual L. delbrueckii strains.

* Corresponding author. Mailing address: Dipartimento Scientifico e Tecnologico, Facoltà di Scienze MM.FF.NN., Università degli Studi di Verona, Strada Le Grazie, Cà Vignal, 1-37134 Verona, Italy. Phone: 39 045 8098917. Fax: 39 045 8098929. E-mail: dellaglio{at}sci.univr.it.

Applied and Environmental Microbiology, October 1999, p. 4351-4356, Vol. 65, No. 10
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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