title: A novel multiplex PCR/RFLP assay for the identification of Streptococcus bovis/Streptococcus equinus complex members from dairy microbial communities based on the 16S rRNA gene.
abstract: The Streptococcus bovis/Streptococcus equinus complex (SBSEC) comprises pathogenic species associated with different degrees with human infections but also spontaneously fermented dairy products. We aimed therefore at developing a specific identification assay for the SBSEC targeting the 16S rRNA gene comprising a multiplex PCR followed by a differentiating restriction fragment length polymorphisms (RFLP). The multiplex PCR assay was positively applied on 200 SBSEC isolates including reference strains. The assay did not yield false-positive amplifications with strains of closely related bacteria and isolates of non-SBSEC streptococci, lactococci, enterococci, and other genera of dairy origin. The downstream RFLP using MseI and XbaI enabled further discrimination of Streptococcus infantarius/S. bovis (biotype II.1) from Streptococcus gallolyticus (biotype I and II.2)/Streptococcus alactolyticus and S. equinus. Furthermore, the newly developed primers can be used directly for Sanger sequencing. Conclusively, this novel PCR/RFLP assay is applicable in the complex dairy microbial communities and provides an important tool to assess the prevalence of members of the SBSEC in dairy products.
authors.affiliation: Laboratory of Food Biotechnology, Institute of Food, Nutrition and Health, ETH Zurich, Switzerland.
authors.forename: Christoph
authors.initials: C
authors.lastname: Jans
authors.forename: Christophe
authors.initials: C
authors.lastname: Lacroix
authors.forename: Leo
authors.initials: L
authors.lastname: Meile
mesh_terms: D001431:Bacteriological Techniques / Q000379:methods*; D005516:Food Microbiology; D020459:Genes, rRNA / Q000235:genetics; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D012680:Sensitivity and Specificity; D013291:Streptococcus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D012336:RNA, Ribosomal, 16S