Dna polymerase extension assay for influenza virus endonuclease

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) C12Q 1/70 (2006.01)

Patent

CA 2222753

An assay for the influenza virus endonuclease has been developed which involves DNA polymerase-catalyzed extension of the viral endonuclease cleavage product using labeled nucleotides and a DNA template containing a 3' region complementary to the product joined to a 5' region consisting of repeated residues. The DNA polymerase coupled assay does not involve gel electrophoretic separation and is amenable to high volume screening of potential inhibitors. Another key feature of the assay is that it is sensitive enoughto detect 200 attomoles of product.

Procédé de détection de l'endonucléase du virus de la grippe consistant en l'extension de l'ADN catalysée par polymérase du produit de clivage de l'endonucléase virale au moyen de nucléotides marqués et d'une matrice d'ADN contenant une région 3' complémentaire du produit réunie à une région 5' constituée par des résidus répétés. Ce procédé utilisant l'ADN polymérase ne nécessite pas de séparation par électrophorèse sur gel et permet d'obtenir un criblage d'un volume important d'inhibiteurs potentiels. Une autre caractéristique clé du procédé est qu'il présente une sensibilité suffisante pour détecter 200 attomoles de produit.

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