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Nouvelles technologies intégrées d'adressage et de détection des interactions moléculaires pour application de biopuces en diagnostic moléculaire in vitro

Abstract : The diagnosis market increased since the advent of molecular biology. More precise and often faster, the in vitro molecular diagnosis (DIV) is more and more used in medical analyses laboratories. DNA chips technology seems to be a good candidate to answer the market expectations. Indeed, this tool allows making several hundreds of analyses simultaneously. Furthermore, it allows giving a global answer of all the present species in the sample without the need of a culture step. Nevertheless, this technology is not optimized for the market of the DIV. Indeed, its use is complex and too expensive in comparison with the current methods (Pasteur microbiology, PCR, Elisa, etc.). So it is necessary to develop an alternative method to reduce the manufacturing cost and simplify the use of DNA chips. First, the goal of this industrial PhD Cifre supported by the Dendris Company was to complete a new prototype of biomolecules deposition based on soft lithography, allowing multiplexing the deposition of oligonucleotides probes along micro and nanometric patterns.This new technology was compared with the reference technologies. Then, we developed an innovative process of surface co-functionalization. This simple and fast process permits to functionalize the DNA chips with both surface chemistry and probes in a single step and to increase the hybridization signals. The second part of this PhD thesis was to couple this new technology with label-free detection using light diffraction. The main difference with fluorescence-based detection was about probes patterning. Indeed, we needed to generate molecular gratings of nanometric lines to diffract efficiently light from a laser beam. We showed that the absolute diffraction intensity increase with the gratings thickness, which is directly correlated with, probes and targets interactions. The second phase of the project was very ambitious and innovative because we demonstrated the feasibility of this label-free detection. And now we can think that this technology will appear as an alternative method for the diagnosis
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Julie Foncy. Nouvelles technologies intégrées d'adressage et de détection des interactions moléculaires pour application de biopuces en diagnostic moléculaire in vitro. Biotechnologies. INSA de Toulouse, 2013. Français. ⟨NNT : 2013ISAT0043⟩. ⟨tel-02003471⟩

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