Multiplexed quantitative detection of pathogens

C - Chemistry – Metallurgy – 12 – P

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C12P 19/34 (2006.01)

Patent

CA 2629076

The invention allows for the quantitative detection of a plurality of pathogens in a single sample. The method includes the amplification of a sample with a plurality of pathogen-specific primer pairs to generate amplicons of distinct sizes from each of the pathogen specific primer pairs. The method further includes the use of a plurality of competitor polynucleotide targets that correspond to each of the pathogen-specific primer pairs. The competitor polynucleotides are added to the reaction mixture at a known concentration to allow for the quantitation of the amount of pathogen in the sample. The method can be used for monitoring pathogen infection in an individual, preferably an immunocompromised individual.

La présente invention concerne la détection quantitative d~une pluralité d~agents pathogènes dans un échantillon unique. Le procédé comprend l~amplification d~un échantillon à l~aide d~une pluralité de paires d~amorces spécifiques à un agent pathogène afin de générer des amplicons de tailles différentes à partir de chacune des paires d~amorces spécifiques à un agent pathogène. Le procédé comprend en outre l~utilisation d~une pluralité de cibles polynucléotidiques concurrentes qui correspondent à chacune des paires d~amorces spécifiques à un agent pathogène. Les polynucléotides concurrents sont ajoutés au mélange réactionnel en une concentration connue afin de permettre la quantification de la quantité d~agent pathogène de l~échantillon. Le procédé peut être utilisé pour surveiller une infection pathogène chez un individu, de préférence chez un immunocompromis.

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