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Système microfluidique µLAS pour l’analyse de l’ADN résiduel : Application au diagnostic de la maladie de Huntington et à l'analyse de l'ADN circulant dans le sang

Rémi Malbec 1
1 LAAS-MICA - Équipe MICrosystèmes d'Analyse
LAAS - Laboratoire d'analyse et d'architecture des systèmes
Abstract : DNA fragments are circulating in the bloodstream. Circulating DNA fragments size, concentration or sequence are analytical information for the clinician or the molecular biology specialists. For instance, circulating DNA, stem from tumoral cells, can serve as biomarkers for cancer detection and follow up. Collecting those information may be difficult for samples presenting minute amount of DNA. In practice, detecting and analyzing residual DNA, with the relevant level of sensitivity, ask for the development and the association of analytical technologies, such as electrophoresis, to molecular biology techniques, such as PCR amplification. In the prospect of simplifying and speeding up the processes, we have developed and optimized µLAS, a microfluidic system for the simultaneous concentration, separation and detection of residual DNA. Furthermore, µLAS has been applied to the diagnostic of Huntington’s disease and the analysis of residual DNA circulating in the bloodstream. Huntington’s disease, is caused by the expansion of CAG/CTG repeats on the Huntingtin gene, and provoke neurological degeneration. The diagnostic of Huntington’s disease consist in amplifying and measuring this expansion. As the amplification of trinucleotide repeat is far from reliable, we benefit from µLAS sensitivity to reduce the number of amplification cycles, and the time to result. Additionally, for the sensitive analysis of circulating DNA by µLAS, we have proposed an original approach, aiming to reduce the blood sample pre-analytical steps to a simple enzymatic digestion followed by a centrifugation step. Finally, the development of a function for the detection of specific sequences has been made by the selective concentration of a target of interest hybridized to a probe. This approach which use some probes fluorescently labelled in volume, has been patented.
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https://tel.archives-ouvertes.fr/tel-01744797
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Submitted on : Tuesday, March 27, 2018 - 4:27:08 PM
Last modification on : Thursday, June 10, 2021 - 3:03:03 AM
Long-term archiving on: : Thursday, September 13, 2018 - 7:37:27 AM

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  • HAL Id : tel-01744797, version 1

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Rémi Malbec. Système microfluidique µLAS pour l’analyse de l’ADN résiduel : Application au diagnostic de la maladie de Huntington et à l'analyse de l'ADN circulant dans le sang. Micro et nanotechnologies/Microélectronique. INSA de Toulouse, 2018. Français. ⟨NNT : 2018ISAT0001⟩. ⟨tel-01744797⟩

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