Viability determination of Helicobacter pylori using propidium monoazide quantitative PCR.

Gemma Agustí, Francesc Codony, Mariana Fittipaldi, Bárbara Adrados, Jordi Morató
Author Information
  1. Gemma Agustí: Laboratori de Microbiologia Sanitària i Mediambiental (MSM Lab)-Aquasost, UNESCO Chair in Sustainability, Universitat Politècnica de Catalunya, 08222 Terrassa, Barcelona, Spain.

Abstract

BACKGROUND: While Helicobacter pylori exists in a bacillary form in both the natural habitat and the human host, detrimental environmental circumstances have been observed to lead to the conversion of H. pylori from the bacillary to the coccoid form. However, the viability or nonviability of coccoid forms remains to be established in H. pylori. The aim of this study was to determine whether the quantitative PCR combined with propidium monoazide could be an alternative and good technique to determine H. pylori viability in environmental samples and, to contribute to understanding of the role of the H. pylori forms.
MATERIALS AND METHODS: Viability, morphological distribution, and the number of live H. pylori cells were determined using a propidium monoazide-based quantitative PCR method, at various time points.
RESULTS: Under adverse environmental conditions was observed the conversion of H. pylori from the bacillary to the coccoid form, and the decrease in amplification signal, in samples that were treated with propidium monoazide, over the time.
CONCLUSIONS: Incorporation of propidium monoazide indicates that there is an increase in H. pylori cells with the damaged membrane over the study, leading to the manifestation of cellular degeneration and death. Consequently, quantitative PCR combined with propidium monoazide contributes to our understanding of the role of H. pylori cells, under adverse environmental conditions.

MeSH Term

Azides
Bacteriological Techniques
Environmental Microbiology
Enzyme Inhibitors
Helicobacter pylori
Humans
Microbial Viability
Microscopy
Polymerase Chain Reaction
Propidium

Chemicals

Azides
Enzyme Inhibitors
propidium monoazide
Propidium

Word Cloud

Created with Highcharts 10.0.0pyloriHpropidiummonoazideenvironmentalquantitativePCRbacillaryformcoccoidcellsHelicobacterobservedconversionviabilityformsstudydeterminecombinedsamplesunderstandingroleViabilityusingtimeadverseconditionsBACKGROUND:existsnaturalhabitathumanhostdetrimentalcircumstancesleadHowevernonviabilityremainsestablishedaimwhetheralternativegoodtechniquecontributeMATERIALSANDMETHODS:morphologicaldistributionnumberlivedeterminedmonoazide-basedmethodvariouspointsRESULTS:decreaseamplificationsignaltreatedCONCLUSIONS:IncorporationindicatesincreasedamagedmembraneleadingmanifestationcellulardegenerationdeathConsequentlycontributesdetermination

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