Skip to Main content Skip to Navigation
Journal articles

Construction of a multiple fluorescence labelling system for use in co-invasion studies of Listeria monocytogenes.

Abstract : BACKGROUND: Existing virulence models are often difficult to apply for quantitative comparison of invasion potentials of Listeria monocytogenes. Well-to-well variation between cell-line based in vitro assays is practically unavoidable, and variation between individual animals is the cause of large deviations in the observed capacity for infection when animal models are used.One way to circumvent this problem is to carry out virulence studies as competition assays between 2 or more strains. This, however, requires invasion-neutral markers that enable easy discrimination between the different strains. RESULTS: A fluorescent marker system, allowing visualization and identification of single L. monocytogenes cells as well as colonies in a non-destructive manner, was developed. Five different fluorescent labels are available, and allowed simultaneous visual discrimination between three differently labelled strains at the single cell level by use of fluorescence microscopy. More than 90% of the L. monocytogenes host cells maintained the fluorescence tags for 40 generations.The fluorescence tags did not alter the invasive capacity of the L. monocytogenes cells in a traditional Caco-2 cell invasion assay, and visual discrimination between invaded bacteria carrying different fluorescent labels inside the cells was possible. CONCLUSION: The constructed fluorescent marker system is stable, easy to use, does not affect the virulence of L. monocytogenes in Caco-2 cell assays, and allows discrimination between differently labelled bacteria after internalization in these cells.
Document type :
Journal articles
Complete list of metadatas

Cited literature [26 references]  Display  Hide  Download

https://www.hal.inserm.fr/inserm-00122150
Contributor : Delphine Autard <>
Submitted on : Wednesday, December 27, 2006 - 1:42:12 PM
Last modification on : Friday, March 27, 2020 - 2:21:56 AM
Long-term archiving on: : Friday, September 21, 2012 - 9:51:22 AM

Identifiers

Collections

Citation

Jens Andersen, Bent Roldgaard, Ariel Lindner, Bjarke Christensen, Tine Licht. Construction of a multiple fluorescence labelling system for use in co-invasion studies of Listeria monocytogenes.. BMC Microbiology, BioMed Central, 2006, 6, pp.86. ⟨10.1186/1471-2180-6-86⟩. ⟨inserm-00122150⟩

Share

Metrics

Record views

437

Files downloads

930