Logo du site
  • English
  • Français
  • Se connecter
Logo du site
  • English
  • Français
  • Se connecter
  1. Accueil
  2. Université de Neuchâtel
  3. Publications
  4. Quantification of Endospore-Forming <i>Firmicutes</i> by Quantitative PCR with the Functional Gene <i>spo0A</i>
 
  • Details
Options
Vignette d'image

Quantification of Endospore-Forming <i>Firmicutes</i> by Quantitative PCR with the Functional Gene <i>spo0A</i>

Auteur(s)
Bueche, Matthieu 
Institut de biologie 
Editeur(s)
Wunderlin, Tina
Roussel-Delif, Ludovic 
Institut de biologie 
Junier, Thomas 
Institut de biologie 
Sauvain, Loïc 
Institut de biologie 
Jeanneret, Nicole 
Institut de biologie 
Junier, Pilar 
Institut de biologie 
In
Applied and Environmental Microbiology, American Society for Microbiology, 2013/79/17/5302-5312
Résumé
Bacterial endospores are highly specialized cellular forms that allow endospore-forming <i>Firmicutes</i> (EFF) to tolerate harsh environmental conditions. EFF are considered ubiquitous in natural environments, in particular, those subjected to stress conditions. In addition to natural habitats, EFF are often the cause of contamination problems in anthropogenic environments, such as industrial production plants or hospitals. It is therefore desirable to assess their prevalence in environmental and industrial fields. To this end, a high-sensitivity detection method is still needed. The aim of this study was to develop and evaluate an approach based on quantitative PCR (qPCR). For this, the suitability of functional genes specific for and common to all EFF were evaluated. Seven genes were considered, but only <i>spo0A</i> was retained to identify conserved regions for qPCR primer design. An approach based on multivariate analysis was developed for primer design. Two primer sets were obtained and evaluated with 16 pure cultures, including representatives of the genera <i>Bacillus</i>, <i>Paenibacillus</i>, <i>Brevibacillus</i>, <i>Geobacillus</i>, <i>Alicyclobacillus</i>, <i>Sulfobacillus</i>, <i>Clostridium</i>, and <i>Desulfotomaculum</i>, as well as with environmental samples. The primer sets developed gave a reliable quantification when tested on laboratory strains, with the exception of <i>Sulfobacillus</i> and <i>Desulfotomaculum</i>. A test using sediment samples with a diverse EFF community also gave a reliable quantification compared to 16S rRNA gene pyrosequencing. A detection limit of about 10<sup>4</sup> cells (or spores) per gram of initial material was calculated, indicating this method has a promising potential for the detection of EFF over a wide range of applications.
URI
https://libra.unine.ch/handle/123456789/8405
DOI
10.1128/AEM.01376-13
Autre version
http://dx.doi.org/10.1128/AEM.01376-13
Type de publication
Resource Types::text::journal::journal article
Dossier(s) à télécharger
 main article: Bueche_M.-Quantification_of_endospore-forming-20140523.pdf (2.32 MB)
google-scholar
Présentation du portailGuide d'utilisationStratégie Open AccessDirective Open Access La recherche à l'UniNE Open Access ORCID

Adresse:
UniNE, Service information scientifique & bibliothèques
Rue Emile-Argand 11
2000 Neuchâtel

Construit avec Logiciel DSpace-CRIS Maintenu et optimiser par 4Sciences

  • Paramètres des témoins de connexion
  • Politique de protection de la vie privée
  • Licence de l'utilisateur final