Nucleic acids as viability markers for bacteria detection using molecular tools
Authors: Cenciarini-Borde, Claire1; Courtois, Sophie; La Scola, Bernard
Source: Future Microbiology, Volume 4, Number 1, February 2009 , pp. 45-64(20)
Publisher: Future Medicine
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Abstract:
A large set of nucleic acid detection methods with good sensitivity and specificity are now available for the detection of pathogens in clinical, food and environmental samples. Given increasing demand, many efforts have been made to combine these methods to assess viability. Genomic DNA PCR amplification has been shown to be inappropriate for distinguishing viable from dead bacteria owing to DNA stability. Many authors have tried to bypass this difficulty by switching to RNA amplification methods such as reverse transcription-PCR and nucleic acid sequence-based amplification. More recently, researchers have developed methods combining specific sample pretreatment with nucleic acid detection methods, notably ethidium or propidium monoazide pretreatment coupled with PCR DNA detection or direct viable count methods and subsequent fluorescent in situ hybridization of 16S rRNA. This review evaluates the performance of these different methods for viability assessment.Keywords: bacteria; direct viable count; ethidium monoazide-PCR; fluorescent in situ hybridization; nucleic acid-based sequence amplification; propidium monoazide-PCR; reverse transcription-PCR; viability assessment; viable but nonculturable
Document Type: Research article
DOI: 10.2217/17460913.4.1.45
Affiliations: 1: CIRSEE (Centre International de Recherche Sur l'Eau et l'Environnement) - Suez Environment, 38 Rue Du Président Wilson 78230 Le Pecq, France and, URMITE, CNRS-IRD UMR 6236, Université de la Méditerranée, Faculté de Médecine, 27 Bd Jean Moulin, 13385 Marseille Cedex 05, France., Email: claire.cen@yahoo.fr
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