Rapid Differentiation and in Situ Detection of 16 Sourdough Lactobacillus Species by Multiplex PCR

Overview

Journal Appl Environ Microbiol

Specialties Environmental Health
Microbiology

Date 2005 Jun 4

PMID 15933001

Citations 6

Authors

Luca Settanni

Douwe Van Sinderen

Jone Rossi

Aldo Corsetti

Affiliations

Soon will be listed here.

Abstract

A two-step multiplex PCR-based method was designed for the rapid detection of 16 species of lactobacilli known to be commonly present in sourdough. The first step of multiplex PCR was developed with a mixture of group-specific primers, while the second step included three multiplex PCR assays with a mixture of species-specific primers. Primers were derived from sequences that specify the 16S rRNA, the 16S-23S rRNA intergenic spacer region, and part of the 23S rRNA gene. The primer pairs designed were shown to exclusively amplify the targeted rrn operon fragment of the corresponding species. Due to the reliability of simultaneously identifying Lactobacillus plantarum, Lactobacillus pentosus, and Lactobacillus paraplantarum, a previously described multiplex PCR method employing recA gene-derived primers was included in the multiplex PCR system. The combination of a newly developed, quick bacterial DNA extraction method from sourdough and this multiplex PCR assay allows the rapid in situ detection of several sourdough-associated lactobacilli, including the recently described species Lactobacillus rossii, and thus represents a very useful alternative to culture-based methodologies.

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