Mise en évidence du caractère infectieux des adénovirus par culture cellulaire couplée à une hybridation in situ

Abstract : Human adenoviruses, affiliated to the Adenoviridae family, are part of the most commonly detected viruses in contaminated surface waters and belong to the 12 priority microorganisms of the contaminant candidate list for drinking water (CCL3, USEPA). The aim of this study was to develop a rapid method for visualization of the infectious status of human adenoviruses by combining cell culture with fluorescent in situ hybridization using a molecular beacon oligonucleotide probe. Fluorescence is generated when the molecular beacon probes undergo a spontaneous conformational change upon formation of a hybrid with homologous mRNA during the viral replication cycle in the host cell. The fluorescent signal is then detectable by imaging methods such as confocal laser scanning microscopy. The experimental protocol (fixation and permeabilization of cells, staining of plasma membranes and nuclei, hybridization conditions) was developed with human adenovirus type 2 as viral model and the host cell line HEK 293A. Despite problems of reproducibility and the need for further optimization of the technique, promising results were obtained with detection of viral mRNA inside the cells in less than 24 hours after infection.
Document type :
Master thesis
File URL :
http://docnum.univ-lorraine.fr/public/BUS_M_2013_BRAGARD_TAMARA.pdf
Complete list of metadatas

Cited literature [93 references]  Display  Hide  Download

https://hal.univ-lorraine.fr/hal-01864511
Contributor : Memoires Ul <>
Submitted on : Thursday, August 30, 2018 - 9:45:00 AM
Last modification on : Thursday, August 30, 2018 - 10:00:56 AM

File

BUS_M_2013_BRAGARD_TAMARA.pdf
Files produced by the author(s)

Identifiers

  • HAL Id : hal-01864511, version 1

Collections

Citation

Tamara Bragard. Mise en évidence du caractère infectieux des adénovirus par culture cellulaire couplée à une hybridation in situ. Microbiologie et Parasitologie. 2013. ⟨hal-01864511⟩

Share

Metrics

Record views

46

Files downloads

6