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Séquençage et PCR à haut débit : application à la détection et la caractérisation d'agents pathogènes respiratoires aviaires et au contrôle de pureté microbiologique des vaccins

Abstract : Detection of pathogens becomes an increasing challenge, since infectious diseases represent major risks for both human and animal health. Globalization of trade and travels, evolution of farming practices and global climatic changes, as well as mass migrations are impacting the biology of pathogens and their emerging potential. This manuscript describes three approaches, based on three innovative technologies of molecular biology applied to the detection of pathogens in three different settings : (i) detection of a list of pathogens using real-time quantitative PCR on a microfluidic platform, (ii) unbiased detection of pathogens in complex matrix, using metagenomics and Illumina (Miseq) sequencing and (iii) genotyping of pathogens without isolation of PCR-enrichment using a 3rd generation NGS (Next Generation Sequencing) platform MinION from Oxford Nanopore Technologies. The three studies shown the contribution of these techniques, each representing distinctive features, suitable for the respective applications. Beyond application of these techniques to the field of microbial diagnostics, their use for the control of veterinary immunological drugs is a priority of this project. Veterinary vaccines are not only submitted to mandatory detection of listed pathogens to be excluded, but also to validation of the genetic identity of vaccine strains. The exponential availability and performances of new PCR or sequencing technologies open cutting-edge perspectives in the field of microbial diagnostic and control.
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Submitted on : Tuesday, April 23, 2019 - 11:08:06 AM
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Guillaume Croville. Séquençage et PCR à haut débit : application à la détection et la caractérisation d'agents pathogènes respiratoires aviaires et au contrôle de pureté microbiologique des vaccins. Biologie moléculaire. Université Paris sciences et lettres, 2017. Français. ⟨NNT : 2017PSLEP028⟩. ⟨tel-02106661⟩

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