G - Physics – 01 – N
Patent
G - Physics
01
N
G01N 33/50 (2006.01) C12Q 1/04 (2006.01) G01N 33/564 (2006.01) G01N 33/68 (2006.01)
Patent
CA 2266124
The invention proposes a method for detecting and/or quantifying, in a biological sample a cytotoxic factor, in particular a gliotoxic factor, with respect to adherent target cells, in particular macroglial cells, the toxicity of which causes the death by apoptosis of said cells. The method consists in providing an initial fraction of said sample, optionally enriched with said toxic factor by previous treatment, incubating said initial toxic factor with a reference culture medium comprising adherent target cells, and detecting and/or quantifying the adherent target cells killed by apoptosis, by flux cytometry, at least one direct or indirect characteristic associated with the apoptotic adherent cells of the whole or part of the incubated medium, which, if it is present and/or is quantified, qualifies the sample as positive, i.e. as containing said toxic factor. The initial biological sample is preferably a urine specimen.
L'invention propose un procédé pour détecter et/ou quantifier, dans un échantillon biologique, un facteur cytotoxique, notamment un facteur gliotoxique, vis-à-vis de cellules adhérentes cibles, notamment les cellules macrogliables, et dont la cytotoxicité induit la mort par apoptose desdites cellules, selon lequel on dispose d'une fraction de départ dudit échantillon, éventuellement enrichie en ledit facteur toxique par un traitement préalable, on incube ladite fraction de départ avec un milieu de culture de référence comprenant des cellules adhérentes cibles, et on détecte et/ou quantifie les cellules adhérentes mortes par apoptose, par cytométric de flux, au moins une caractéristique directe ou indirecte, associée aux cellules adhérentes apoptotiques de tout ou partie du milieu incubé, qui, si elle existe et/ou est quantifiée, qualifie ledit échantillon biologique comme positif, c'est-à-dire contenant ledit facteur toxique. L'échantillon biologique de départ est de préférence un échantillon d'urine.
Malcus-Vocanson Carine
Mandrand Bernard
Perron Herve
Bio Merieux
Ogilvy Renault Llp/s.e.n.c.r.l.,s.r.l.
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