Application of multilocus sequence analysis (MLSA) for rapid identification of Enterococcus species based on rpoA and pheS genes

Naser, Sabri M.; Thompson, Fabiano L.; Hoste, Bart; Gevers, Dirk; Dawyndt, Peter; Vancanneyt, Marc; Swings, Jean

doi:10.1099/mic.0.27840-0

Cited by 367 publications

(322 citation statements)

References 53 publications

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“…The multiplex PCR assay based on sodA gene reported by Jackson, Fedorka-Cray, and Barrett (2004) was applied to confirm species identity. Finally, pheS partial sequence was obtained for strain WFE31 as previously reported (Naser et al, 2005) and compared in GenBank/EMBL/DDBJ database. DNA amplifications were performed by means of T1 Thermocycler (Biometra, Göttingen, Germany).…”

Section: Identification Of Enterococci At Species Level and Strain DImentioning

confidence: 99%

Production, stability, gene sequencing and in situ anti-Listeria activity of mundticin KS expressed by three Enterococcus mundtii strains

Settanni¹,

Guarcello²,

Gaglio³

et al. 2014

Food Control

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a b s t r a c tThree enterococci (WFE3, WFE20 and WFE31) selected as presumptive bacteriocin producers were found to be active against Listeria monocytogenes. In this study, due to their potential industrial/food applications, the three bacterial isolates were extensively characterized. Identification was performed by means of a combined 16S rRNA gene sequencing and multiplex PCR approach, and was confirmed with the sequencing of a partial region of a protein-encoding gene, namely pheS. The three isolates belonged unequivocally to the species Enterococcus mundtii. The randomly amplified polymorphic DNA (RAPD) analysis recognized three distinct strains. The supernatants were mainly active against Listeria spp., but some lactic acid bacteria were also inhibited. The proteinaceous nature of the three supernatants was detected after treatment with proteinase K, protease B and trypsin. The bacteriocins were found to be heat resistant, stable in a large pH range and in presence of ethanol. The bacteriocins were not adsorbed onto the surface of the producer cells and their effect was bactericidal. The production of bacteriocins was higher at neutral pHs and temperatures in the range 30e37 C. The active supernatants did not show cytotoxicity against human erythrocytes and the three strains were susceptible to the action of common antibiotics. The genetic characterization of the bacteriocin genes showed that all three strains produced mundticin KS. They produced it in five food model systems, sterilized by thermal treatment or filtration, prepared from fresh vegetables, cereals, cheeses, meats and fishes. The in situ anti-listerial activities of the strains WFE3, WFE20 and WFE31 were quantitatively different.

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Section: Identification Of Enterococci At Species Level and Strain DImentioning

confidence: 99%

Production, stability, gene sequencing and in situ anti-Listeria activity of mundticin KS expressed by three Enterococcus mundtii strains

Settanni¹,

Guarcello²,

Gaglio³

et al. 2014

Food Control

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“…The 16S rRNA gene was amplified using the primer pair pA (59-AGAGTTTGATCCTGGCTCAG-39) and pH (59-AAGGAGGTGATCCAGCCGCA-39) (Edwards et al, 1989). The pheS gene was amplified using primers pheS-21-F (59-CAYCCNGCHCGYGAYATGC-39) and pheS-23-R (59-GGRTGRACCATVCCNGCHCC-39) (Naser et al, 2005). The amplified products were purified using the QIAquick PCR Purification kit (Qiagen).…”

mentioning

confidence: 99%

Weissella uvarum sp. nov., isolated from wine grapes

Nisiotou¹,

Dourou²,

Filippousi³

et al. 2014

International Journal of Systematic and Evolutionary Microbiology

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Two bacterial strains (B18BM42 T and B18NM6) were recovered during a study of bacterial diversity on wine grapes (Vitis vinifera L.) from the Nemea region in Greece. Phylogenetic analysis based on 16S rRNA gene sequences placed the two strains within the genus Weissella, and found them to be most closely related to Weissella minor NRIC 1625 T followed by Weissella viridescens NRIC 1536 T (99.1 and 98.9 % sequence similarity, respectively). The level of DNA-DNA relatedness between strains B18NM42 T and W. minor NRIC 1625 T or W. viridescens NRIC 1536 T was 31.9 and 35.0 %, respectively. The two novel strains could be genetically differentiated from their closest relatives by REA-PFGE (restriction enzyme analysis-pulse field gel electrophoresis), RAPD (randomly amplified polymorphic DNA) and rep-PC R analyses (repetitive sequence-based PCR). Physiological examination showed that the novel strains can be distinguished from phylogenetically related species by their ability to grow at 42 6C and by certain carbohydrate fermentations. Based on the evidence above, the affiliation of the two strains to a novel species with the proposed name Weissella uvarum sp. nov. is suggested. The type strain is B18NM42 T (5DSM 28060 T 5NCCB 100484 T ).

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“…The MLSA technique has also been applied for rapid and reliable species identification in other bacterial genera, e.g. Enterococcus, Aeromonas and Haemophilus (Naser et al, 2005;Nørskov-Lauritsen et al, 2004;Soler et al, 2004). …”

Section: Resultsmentioning

confidence: 99%

Multilocus sequence analysis supports the taxonomic position of Astragalus glycyphyllos symbionts based on DNA–DNA hybridization

Gnat

Małek

Oleńska

et al. 2016

International Journal of Systematic and Evolutionary Microbiology

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In this study, the phylogenetic relationship and taxonomic status of six strains, representing different phenons and genomic groups of Astragalus glycyphyllos symbionts, originating from Poland, were established by comparative analysis of five concatenated housekeeping gene sequences (atpD, dnaK, glnA, recA and rpoB), DNA-DNA hybridization and total DNA G+C content. Maximum-likelihood phylogenetic analysis of combined atpD, dnaK, glnA, recA and rpoB sequence data placed the studied bacteria into the clade comprising the genus Mesorhizobium. In the core gene phylograms, four A. glycyphyllos nodule isolates (AG1, AG7, AG15 and AG27) formed a cluster common with Mesorhizobium ciceri, whereas the two other A. glycyphyllos symbionts (AG17 and AG22) were grouped together with Mesorhizobium amorphae and M. septentrionale. The species position of the studied bacteria was clarified by DNA-DNA hybridization. The DNA-DNA relatedness between isolates AG1, AG7, AG15 and AG27 and reference strain M. ciceri USDA 3383 T was 76.4-84.2 %, and all these A. glycyphyllos nodulators were defined as members of the genomospecies M. ciceri. DNA-DNA relatedness for isolates AG17 and AG22 and the reference strain M. amorphae ICMP 15022 T was 77.5 and 80.1 %, respectively. We propose that the nodule isolates AG17 and AG22 belong to the genomic species M. amorphae. Additionally, it was found that the total DNA G+C content of the six test A. glycyphyllos symbionts was 59.4-62.1 mol%, within the range for species of the genus Mesorhizobium.

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Application of multilocus sequence analysis (MLSA) for rapid identification of Enterococcus species based on rpoA and pheS genes

Cited by 367 publications

References 53 publications

Production, stability, gene sequencing and in situ anti-Listeria activity of mundticin KS expressed by three Enterococcus mundtii strains

Production, stability, gene sequencing and in situ anti-Listeria activity of mundticin KS expressed by three Enterococcus mundtii strains

Weissella uvarum sp. nov., isolated from wine grapes

Multilocus sequence analysis supports the taxonomic position of Astragalus glycyphyllos symbionts based on DNA–DNA hybridization

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