Method for quantifying a ratio between at least two nucleic...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) C12P 19/34 (2006.01)

Patent

CA 2448769

The invention provides a method for quantifying an initial ratio of the amounts of at least two nucleic acids of interest in a sample by means of a multiplex nucleic acid amplification reaction, comprising: - amplifying said nucleic acids of interest in said amplification reaction; - measuring the amount of at least two nucleic acids of interest at at least two different time points in said reaction; - determining from at least two of said measurements the amplification rate of said at least two nucleic acids of interest; - comparing said rates with a reference; and - determining from said comparison said initial ratio of the amounts of said at least two nucleic acids of interest in said sample. Preferably, at least one variable factor in said nucleic acid amplification reaction is adjusted in order to allow detectable levels of all nucleic acids of interest to be reached before an amplification and/or detection limit of one or more of the nucleic acids of interest is reached. With a method of the invention, small differences between ratios can be determined. Moreover, direct measurement of the initial nucleic acid ratio can be determined. A method of the invention is for instance suitable for determining functioning of a cellular organism, for determining the staging of a disease, and for determining therapeutic activity and/or possible side-effects of a medicament or candidate compound.

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