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Système de biopuce optique en temps réel: application au diagnostic génétique

Nathalie Bassil 1
1 Laboratoire Charles Fabry de l'Institut d'Optique / Macsybio - bio
LCFIO - Laboratoire Charles Fabry de l'Institut d'Optique
Abstract : In the second half of the twentieth century, the world witnessed the birth of new technologies such as DNA biochips. The use of Surface Plasmon Resonance Imaging in those techniques is emerging nowadays. It allows following without using any marker, in real time and in parallel, the interactions taking place on a metal surface often made of gold. We exploit this imaging technique to analyze DNA/DNA interactions in order to achieve a genetic diagnosis. Therefore, we fixed single strands of biotinylated DNA on gold surfaces via a self-assembled structure composed successively by a mercaptoundecanoic acid, a positively charged polymer and an ExtrAvidin layer. The results obtained when studying DNA/DNA hybridization, show that the system optical response depends on the complementarity of DNA strands. As a matter of fact, we could easily determine the presence of a deletion meanwhile the single base mismatch was difficult to detect. To solve this problem we studied the effect of oligonucleotide's length. The results highlighted the importance of DNA's base composition and melting temperature. After representing the sensor's response as a function of DNA's melting temperature calculated under certain conditions of salt concentration, we were able to predict the hybridization signal for any single strand DNA used and to understand why it was difficult to detect the single base mismatch. Based on our prediction method we designed a set of oligonucléotides that allow the detection of six mutations located on the Cystic Fibrosis gene. The results obtained agreed with the calculated value and opened the way for the realization of a DNA chip for genetic diagnosis.
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https://pastel.archives-ouvertes.fr/tel-00008933
Contributor : Nathalie Bassil <>
Submitted on : Wednesday, May 11, 2005 - 5:52:38 AM
Last modification on : Wednesday, October 14, 2020 - 4:00:12 AM
Long-term archiving on: : Monday, September 20, 2010 - 12:46:12 PM

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  • HAL Id : tel-00008933, version 2

Citation

Nathalie Bassil. Système de biopuce optique en temps réel: application au diagnostic génétique. Physique [physics]. Université Paris Sud - Paris XI, 2005. Français. ⟨tel-00008933v2⟩

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