Detection of microbial metabolites

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/44 (2006.01) C07F 9/572 (2006.01) C12Q 1/04 (2006.01)

Patent

CA 2281709

A method of detecting a phosphatidylinositol-specific phospholipase C enzyme by means of a substrate which is cleaved by said enzyme and yields a dye when the chromophoric portion of the substrate is dimerized and oxidized; the invention teaches using in such method, as a novel substrate, a 3-indoxyl- myoinositol-1-phosphate compound of formula (I) wherein R is selected from the group consisting of hydrogen and C1-4 alkyl, while R1, R2, R3, and R4 are radicals selected from the group consisting of hydrogen and chromogenic substituents, or of a salt of said formula (I) compound. The invention provides for a safe, sensitive and commercially viable detection of potentially pathogenic bacterial activity of such microbes as Bacillus cereus, B. Thuringiensis, Staphylococcus aureus and various Listeria strains in potentially infected materials including physiological samples or consumable goods such as foods and beverages.

L'invention concerne un procédé pour détecter une enzyme phospholipase C spécifique au phosphatidylinositol, à l'aide d'un substrat qui est coupé par cet enzyme et produit un colorant lorsque la partie chromophore du substrat est dimérisée et oxydée. L'invention a également pour objet l'utilisation, avec ce procédé, d'un nouveau substrat, un composé 3-indoxyle-myo-inositol-1-phosphate selon la formule (I). Dans cette dernière, R est sélectionné dans le groupe comportant de l'hydrogène et de l'alkyle C¿1?-¿4?, et R¿1?, R¿2?, R¿3? et R¿4? sont des radicaux sélectionnés dans le groupe se composant d'hydrogène et de substituants chromogènes, ou d'un sel de ce composé selon la formule (I). L'invention a pour objet un système, sans danger; présentant une grande sécurité, et viable d'un point de vue commercial, permettant de détecter une activité bactérienne potentiellement pathogène de ces microbes, tels que le Bacillus cereus, B. Thuringiensis, Staphylococcus aureus et diverses souches de Listeria dans des matériaux potentiellement infectés comprenant des échantillons physiologiques ou des marchandises consommables telles que des aliments et des boissons.

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