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Master Thesis

Mise en place d’une prestation en routine au sein d’un CRB : génotypage par l’étude de SNP d’une collection biologique issue de la recherche clinique

Abstract : Biobanks have been created and defined in order to recollect with high controlled procedures samples from clinical research protocol. These organizations are part of rigorous quality process required by NF S96-900 and ISO 9001 norms. They allow the research field and industrial to get access to high standard and tracked resources. In this context, in 2005 CRB HUEP-UPMC recollected human blood samples from a clinical protocol in order to demonstrate a link between specific gene polymorphism and the morbidity/mortality of patients included in this project. To answer to their objective, a genotyping method which is Single Nucleotide Polymorphisms (SNP) technique has been selected and should be validated by the CRB. This new analysis process is considered as a routine performance which required specific conditions for the handling samples according to their origins; DNA extracted by the CRB HUEP-UPMC or by the user himself. The development of the technique allowed to genotype 1929 patients and to validate the SNP genotyping method. Several blood samples storage conditions, DNA extraction and storage conditions were tested and analyzed by DNA quality controls. The results demonstrated that genotyping is reliable if DNA is compliant (meaning DNA concentration >50 ng/μl, 1.7< DO 260/280 nm <2, DO 260/230 nm >1.8), regardless of blood conservation and DNA extraction methods. By contrast, DNA storage process is the most critical step which requires to be conditioned in cryovial and stored at -20°C in the long term. This information is crucial to provide quality service to external users even for which we have no view of the pre-analytical phase.
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Master Thesis
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https://dumas.ccsd.cnrs.fr/dumas-01543318
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Submitted on : Tuesday, June 20, 2017 - 4:21:56 PM
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Catherine Delmau-Laffargue. Mise en place d’une prestation en routine au sein d’un CRB : génotypage par l’étude de SNP d’une collection biologique issue de la recherche clinique. Biologie cellulaire. 2015. ⟨dumas-01543318⟩

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