Methods and compositions for detection of segmented negative...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/70 (2006.01) C12Q 1/02 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2544847

The present invention provides compositions and methods for the detection of the presence, absence, or quantity of a segmented negative strand RNA virus such as an influenza virus. A genetically engineered vertebrate cell comprising an artificial segment comprising a 5' UTR and a 3' UTR of a segmented negative strand RNA virus and an open reading frame of a reporter gene, preferably in an anti-sense orientation, is contacted with a biological specimen suspected of comprising a segmented negative strand virus. Infection of the cell with a segmented negative strand RNA virus results in expression of a polypeptide encoded by the reporter gene. A genetically engineered cell of the invention can also comprise a recombinant DNA encoding the artificial segment. The recombinant DNA can comprise a promoter for RNA Polymerase I for directing transcription of the artificial segment.

La présente invention concerne des compositions et des méthodes de détection de la présence, de l'absence, ou de la quantité d'un virus à ARN à brin négatif segmenté tel que le virus de la grippe. Une cellule vertébrée obtenue par génie génétique et comprenant un segment artificiel contenant 5' UTR et 3' UTR d'un virus à ARN à brin négatif segmenté et une phase de lecture ouverte d'un gène reporter, de préférence dans une orientation anti-sens, est mise en contact avec un spécimen biologique susceptible de contenir un virus à brin négatif segmenté. L'infection de la cellule par un virus à ARN à brin négatif segmenté entraîne l'expression d'un polypeptide codé par le gène reporter. La cellule obtenue par génie génétique de l'invention peut également comprendre un ADN de recombinaison codant pour le segment artificiel. L'ADN de recombinaison peut comprendre un agent promoteur d'ARN polymérase I dirigeant la transcription du segment artificiel.

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