Method of analyzing a target nucleic acid

A method of analyzing a target nucleic acid by applying a nucleic acid amplification reaction to a test solution containing the target nucleic acid, wherein an amplified product is labeled with a marker molecule, said method comprises:

• (a) performing a nucleic acid amplification reaction of the target nucleic acid in a test solution containing a forward primer and a reverse primer, a substrate comprising nucleotides, a nucleic acid polymerase and a target nucleic acid, wherein the forward primer is in a lower amount than the reverse primer or the reverse primer is in a lower amount than the forward primer, and the primer present in a lower amount is labeled with a marker molecule capable of generating a detectable signal to form a labeled primer, the nucleic acid amplification reaction being performed until the primer present in a lower amount is consumed;
• (b) measuring a signal from the marker molecule in the test solution after initiation of the nucleic acid amplification reaction;
• (c) evaluating a fluctuation motion of the labeled primer and the amplified nucleic acid which is labeled with the marker molecule, in the test solution on the basis of the signal detected;
• (d) determining a number of cycles of the nucleic acid amplification reaction performed until the labeled primer has been completely consumed, and a yield of the amplified nucleic acid which is labeled with the marker molecule, based on an evaluation result of the step (c);
• and
• (e) quantifying an initial amount of the target nucleic acid on the basis of the number of cycles of the nucleic acid amplification reaction and the yield of the amplified nucleic acid which is labeled with the marker molecule.