DVC-FISH and PMA-qPCR techniques to assess the survival of Helicobacter pylori inside Acanthamoeba castellanii

Research in Microbiology
Laura Moreno-MesoneroMaria A Ferrús

Abstract

Free-living amoebae (FLA) are ubiquitous microorganisms commonly found in water. They can act as Trojan Horses for some amoeba-resistant bacteria (ARB). Helicobacter pylori is a pathogenic bacteria, suggested to be transmitted through water, which could belong to the ARB group. In this work, a co-culture assay of H. pylori and Acanthamoeba castellanii, one of the most common FLA, was carried out to identify the presence and survival of viable and potentially infective forms of the bacteria internalized by the amoeba. Molecular techniques including FISH, DVC-FISH, qPCR and PMA-qPCR were used to detect the presence of internalized and viable H. pylori. After 24 h in co-culture and disinfection treatment to kill extra-amoebic bacteria, viable H. pylori cells were observed inside A. castellanii. When PMA-qPCR was applied to the co-culture samples, only DNA from internalized H. pylori cells was detected, whereas qPCR amplified total DNA from the sample. By the combined DVC-FISH method, the viability of H. pylori cells in A. castellanii was observed. Both specific techniques provided evidence, for the first time, that the pathogen is able to survive chlorination treatment in occurrence with A. castellanii and could be very useful met...Continue Reading

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Citations

Mar 8, 2019·Microbiology·Shravanthi S Kumar, Asit Ranjan Ghosh
Nov 21, 2018·Foodborne Pathogens and Disease·Maria Consuelo Pina-PérezMaria Antonia Ferrús
May 11, 2018·Microbiology and Molecular Biology Reviews : MMBR·Skander HathroubiKaren M Ottemann
Jun 25, 2020·Pathogens·Kimberly Sánchez-AlonzoApolinaria García-Cancino
Mar 25, 2019·Experimental Parasitology·Rasha Maal-BaredDiana Axelsson-Olsson
Jun 20, 2021·Folia Microbiologica·Mohammad Ridwane MungrooNaveed Ahmed Khan
Jan 11, 2020·The Science of the Total Environment·Marta RusiñolRosina Girones

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