C - Chemistry – Metallurgy – 12 – Q
Patent
C - Chemistry, Metallurgy
12
Q
C12Q 1/68 (2006.01) C07H 21/04 (2006.01) C12P 19/34 (2006.01)
Patent
CA 2489373
A real-time PCR method for detecting and quantifying the presence or absence of Renibacterium salmoninarum, the causative organism of Bacterial Kidney Disease, in a sample is disclosed. The method includes the steps of combining with a sample a pair of PCR primers that bind to a DNA that is specific for the organism and a labeled probe that also binds to the DNA, and performing one or more rounds of PCR on the sample. The probe undergoes a change with each round of amplification of the DNA during the PCR reaction, which change causes a change in the signal provided by the label, thus providing a detection of the DNA and, if desired, a quantification, of the level of the DNA, and thus the level of the organism, in the sample. Figure 1 is a diagrammatic representation showing the placement of the preferred primers and probe of the invention on the p57 gene in relation to the primers used for nested PCR.
L'invention concerne un proc~d~ PCR (r~action en cha¹ne de la polym~rase) en temps r~el pour d~tecter et quantifier la pr~sence ou l'absence dans un ~chantillon de Renibacterium salmoninarum, l'organisme provoquant la maladie r~nale bact~rienne. Le proc~d~ selon l'invention comprend les ~tapes suivantes : combinaison avec un ~chantillon d'une paire d'amorces PCR qui se lient ~ un ADN sp~cifique de l'organisme et d'une sonde marqu~e qui se lie ~galement ~ l'ADN, et ex~cution d'un ou de plusieurs cycles de r~action PCR sur l'~chantillon. La sonde subit une modification ~ chaque cycle d'amplification de l'ADN pendant la r~action PCR, cette modification provoquant une variation dans le signal fourni par le marqueur, permettant ainsi une d~tection de l'ADN et, si d~sir~, une quantification du niveau de l'ADN et donc du niveau de l'organisme dans l'~chantillon.
Overturf Kenneth E.
Powell Madison S.
Idaho Research Foundation
Oyen Wiggs Green & Mutala Llp
The United States Of America As Represented By The Secretary Of
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