The spxB gene as a target to identify Lactobacillus casei group species in cheese

Sardaro, Maria Luisa Savo; Levante, Alessia; Bernini, Valentina; Gatti, Monica; Neviani, Erasmo; Lazzi, Camilla

doi:10.1016/j.fm.2016.05.004

Cited by 18 publications

(12 citation statements)

References 28 publications

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“…Obtaining such results for new isolates could lead to their incorrect classification. It should also be noted that our mean Tm values for individual species differed significantly from the analogous results obtained by Sardaro et al [40]. Such an observation puts a big question mark over the possibility of comparing results obtained from different laboratories using the described procedure.…”

Section: High-resolution Melting Analysiscontrasting

confidence: 64%

“…In this study, two sets of primers, which were developed based on spxB and groEL gene sequences, were used to differentiate three closely related species [39,40]. For the first experimental system (spxB) the following average melting temperatures were obtained: 81.6 ± 0.04 • C for L. casei, 81.3 ± 0.15…”

Section: High-resolution Melting Analysismentioning

confidence: 99%

“…The HRM-PCR reaction was carried out with two set of primers, based on the spxB and groEL genes (Table S1) [39,40]. PCR reactions were performed in 15-µL solutions containing 7.…”

Section: Real-time Pcr and High-resolution Melting Analysismentioning

confidence: 99%

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Molecular Routes to Specific Identification of the Lactobacillus Casei Group at the Species, Subspecies and Strain Level

Jarocki

Komoń-Janczara

Glibowska

et al. 2020

IJMS

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The genus Lactobacillus includes, among others, Lactobacillus casei, Lactobacillus paracasei and Lactobacillus rhamnosus, species that are collectively referred to as the Lactobacillus casei group. Many studies have shown that strains belonging to this group may decrease lactose intolerance, the effects of inflammatory bowel disease, diarrhea, constipation, food allergies and even colon cancer. Moreover, evidences exists of positive effects of these bacteria on mucosal immunity and blood cholesterol level. Because of their beneficial influence on human health, many of them are used as food additives and probiotic pharmaceuticals. It should be stressed that health-promoting properties are not attributed at the species level, but to specific strains. Therefore, procedures are necessary to allow specific identification at each phylogenetic level—genus, species and strain. In this paper we present a practical overview of molecular methods for the identification and differentiation of L. casei bacteria. The research included 30 bacterial strains belonging to three species: L.casei, L. paracasei and L. rhamnosus. Among the tested procedures were genus- and species-specific PCR, multiplex-PCR, Real-Time HRM analysis, RFLP-PCR, rep-PCR, RAPD-PCR, AFLP-PCR, and proteomic methods such as MALDI-TOF MS typing and SDS-PAGE fingerprinting. The obtained results showed that multiplex-PCR and MALDI-TOF MS turned out to be the most useful methods to identify the tested bacteria at the species level. At the strain level, the AFLP-PCR method showed the highest discriminatory power. We hope that the presented results will allow for the easy selection of an appropriate procedure, depending on the experiment conducted and the equipment capabilities of any given laboratory.

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Section: High-resolution Melting Analysiscontrasting

confidence: 64%

Section: High-resolution Melting Analysismentioning

confidence: 99%

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Molecular Routes to Specific Identification of the Lactobacillus Casei Group at the Species, Subspecies and Strain Level

Jarocki

Komoń-Janczara

Glibowska

et al. 2020

IJMS

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“…The ability to discriminate between both Photobacterium subspecies was validated on a total of 19 Phdp and 11 Phdd strains isolated from Europe, USA and Japan, proving the assay is able to overcome the documented limitation due to the genetic variability of isolates from diverse hosts and geographical origins (Costas et al., ; Magarinos, Toranzo, Barja, & Romalde, ). The observed difference of at least 0.3°C between the two subspecies has been already proved sufficient/suitable for differentiating virus and bacterial species of medical and veterinary importance (Gelaye et al., ; Miller, Zorn, & Brielmeier, ) as well as for the monitoring of bacterial communities in food (Sardaro et al., ). In fact, both classical melting curve analysis and High Resolution Melting (HRM) analysis have been described as powerful techniques for variant scanning and genotyping, enabling to analyse even single genetic variants in nucleic acid sequences (Wittwer, ).…”

Section: Discussionmentioning

confidence: 99%

Development of a real‐time PCR assay for rapid detection and quantification of Photobacterium damselae subsp. piscicida in fish tissues

Carraro

Rovere

Ferraresso

et al. 2017

Journal of Fish Diseases

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The availability of a rapid and accurate method for the diagnosis of Photobacterium damselae subsp. piscicida (Phdp), able to discriminate its strictly correlated subsp. damselae (Phdd), formally known as Vibrio damsela, is essential for managing fish pasteurellosis outbreaks in farmed fish. A single-step, high-sensitivity real-time PCR assay for simultaneous detection and quantification of P. damselae was designed targeting partial of the sequence of the bamB gene and tested for specificity and sensitivity on laboratory-generated samples as well as on experimentally infected seabream tissue samples. With a limit of detection (LOD) of one copy in pure bacterial DNA, the sensitivity was higher than all methods previously reported. Validation in target and non-target bacterial species proved the assay was able to discriminate Phdd-Phdp subspecies from diverse hosts/geographical origins and between non-target species. In addition, two SNPs in the target amplicon region determine two distinctive qPCR dissociation curves distinguishing between Phdp-Phdd. This is the first time that a molecular method for P. damselae diagnosis combines detection, quantification and subspecies identification in one step. The assay holds the potential to improve the knowledge of infection dynamics and the development of better strategies to control an important fish disease.

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“…Lactobacillus casei is widely distributed in a large number of habitats, being isolated from multiple food products (1). Since it is genetically well adapted to the gastrointestinal and reproductive tract environment of humans and animals (2), several L. casei strains are used as probiotics in functional foods (3, 4).…”

Section: Genome Announcementmentioning

confidence: 99%

High-Quality Draft Genome Sequence of Lactobacillus casei Strain Z11, Isolated from a Human Adult Intestinal Biopsy Sample

et al. 2017

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The spxB gene as a target to identify Lactobacillus casei group species in cheese

Cited by 18 publications

References 28 publications

Molecular Routes to Specific Identification of the Lactobacillus Casei Group at the Species, Subspecies and Strain Level

Molecular Routes to Specific Identification of the Lactobacillus Casei Group at the Species, Subspecies and Strain Level

Development of a real‐time PCR assay for rapid detection and quantification of Photobacterium damselae subsp. piscicida in fish tissues

High-Quality Draft Genome Sequence of Lactobacillus casei Strain Z11, Isolated from a Human Adult Intestinal Biopsy Sample

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