Nucleic acid quantitation by co-amplification of target with multiple

Abstract

The present invention is related to an improved method for the quantification of nucleic acid, which can be performed with a minimal amount of nucleic acid amplification reactions. The method according to the invention for the quantification of analyte nucleic acid in a sample comprises the steps of: adding to the sample different respective amounts of different nucleic acid constructs, each construct being distinguishable from the analyte nucleic acid and capable of being co-amplified with the analyte nucleic acid; subjecting the sample to a nucleic acid amplification procedure, using amplification reagents capable of reacting with both the analyte nucleic acid and the nucleic acid constructs; detecting the relative amounts of amplificates derived from analyte nucleic acid and each nucleic acid construct; calculating the amount of analyte nucleic acid from said relative amounts. Each nucleic acid construct is different; the nucleic acid constructs can be distinguished from one another and from the analyte nuclic acid. The nucleic acid constructs do resemble each other, and the analyte nucleic acid, in that all are capable of reacting with the same amplification reagents.