Measuring viral reverse transcriptase activity

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/70 (2006.01)

Patent

CA 2379488

In clinical settings as well as in a drug-discovery context, the impact of an agent that may affect reverse transcriptase (RT) can be measured and even quantified by bringing a sample, which may contain an RT, into contact with an RNA template, a primer complementary to the RNA template, and appropriate oligonucleotide-specific primers, under conditions such that they react, in the presence of RT, to form a cDNA product in inverse proportion to the effect of the agent. The amount of any resultant cDNA product then can be measured. The approach is readily implemented as a real-time, quantitative kinetic assay for RT activity.

Dans les milieux cliniques et dans le contexte de la découverte de médicaments, l'impact d'un agent susceptible affecter la transcriptase inverse virale (TI) peut être mesuré et même quantifié par mise en contact d'un échantillon contenant la TI avec une matrice ARN, une amorce complémentaire de la matrice ARN et des amorces propres aux oligonucléotides appropriées, dans des conditions telles qu'ils réagissent, en présence de la TI, de manière à former un produit d'ADNc dans des proportion inverse à l'effet de l'agent. La quantité de produit d'ADNc obtenue peut alors être mesurée. Ce procédé peut être mis en oeuvre rapidement pour réaliser un dosage cinétique quantitatif en temps réel de l'activité de la transcriptase inverse virale.

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