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Elaboration de biocapteurs électrochimiques d'ADN à base de nano-structure de polypyrrole pour le diagnostic de la tuberculose

Abstract : Tuberculosis is a contagious disease that usually attacks the lungs but can sometimes reach other parts of the body such as the kidneys, ganglia and bones. This disease is responsible for killing nearly 1.8 million people each year worldwide. Therefore, we have an urgent need to put analytical means to detect the DNA of the bacterium Mycobacterium tuberculosis, responsible for the spread of tuberculosis. The research developed in this thesis consists in the development of a diagnostic tool for the detection of genomic DNA of M.Tuberculosis bacteria after lysis and amplification by PCR. In this perspective, we focused on the development of biosensors based on different polypyrrole morphologies as transducers for the electrochemical detection of DNA of the rpob gene of Mycobacterium tuberculosis. A study of the impact of different morphologies on the electrical properties of materials as well as the performance of electrochemical DNA biosensors was carried out. We also studied the effect of the functionalization by different linear and branched chains on the structure of polypyrrole nanomaterials and their effects on the number of immobilized biomolecules. The strategy adopted for the development of these materials is a biosensor construction carried out step by step. The various layers constituting it have been characterized by different surface techniques such as electrochemical and optical techniques, scanning electron microscopy and atomic force electron microscopy. The properties of the biosensors were monitored through the redox properties of the ferrocenyl groups. The electrochemical detection of the target DNA evaluated with these biosensors based on polypyrrole nanostrcutrue compared to the results obtained with a biosensor based on polypyrrole compact layer shows a higher detection sensitivity.This demonstrates a potential for using the high surface area of polypyrrole nanostructures as a transducer and the use of soft modification approaches. The biosensors were applied to the detection of DNA in real samples of genomic DNA of Mycobacterium tuberculosis and mutated DNA with resistance to rifampicin. The biosensors based on polypyrrole nanostructures demonstrate potential application in DNA detection and the ability to discriminate the mutant rpoB gene and could be used as a platform in biosensor technology.
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https://tel.archives-ouvertes.fr/tel-01947556
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Submitted on : Friday, December 7, 2018 - 1:03:06 AM
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Rabih Khoder. Elaboration de biocapteurs électrochimiques d'ADN à base de nano-structure de polypyrrole pour le diagnostic de la tuberculose. Matériaux. Université Paris-Saclay, 2018. Français. ⟨NNT : 2018SACLS094⟩. ⟨tel-01947556⟩

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