Signal extraction for a target nucleic acid sequence

A computer readable storage medium containing instructions to configure a processor to perform a method for extracting a signal for a target nucleic acid sequence from signals for two target nucleic acid sequences comprising a first target nucleic acid sequence and a second target nucleic acid sequence in a sample, the method comprising:

• (a) receiving signals from the sample at a relatively high detection temperature and a relatively low detection temperature;
• wherein the first target nucleic acid sequence in the sample is detected by a first signal-generating means and the second target nucleic acid sequence in the sample is detected by a second signal-generating means;
• wherein the detection is performed at one or more cycles of the signal-generating process to obtain a signal value at each of the one or more cycles;
• and wherein the two signals to be generated by the two signal-generating means are not differentiated by a single type of detector;
• (b) mathematically processing the signal values obtained in the step (a) by:
• using a second initial reference value to extract the signal for the first target nucleic acid sequence, or
• using a first initial reference value to extract the signal for the second target nucleic acid sequence;
• wherein the first initial reference value is a value representing a relationship of change in signals provided by the first signal-generating means at the relatively high detection temperature and the relatively low detection temperature, and the second initial reference value is a value representing a relationship of change in signals provided by the second signal-generating means at the relatively high detection temperature and the relatively low detection temperature;
• and wherein the first initial reference value is predetermined from a separate reaction using the first target nucleic acid sequence and the first signal-generating means and, the second initial reference value is predetermined from a separate reaction using the second target nucleic acid sequence and the second signal-generating means;
• and
• (c) identifying whether the extracted signal satisfies an accuracy criterion;
• when the extracted signal does not satisfy the accuracy criterion, the step (b) is repeated using an amended reference value instead of the initial reference value to re-extract the signal for the first target nucleic acid sequence or the second target nucleic acid sequence;
• wherein the amended reference value is determined such that the re-extracted signal satisfies the accuracy criterion.

A computer program to be stored on a computer readable storage medium to configure a processor to perform a method for extracting a signal for a target nucleic acid sequence from signals for two target nucleic acid sequences comprising a first target nucleic acid sequence and a second target nucleic acid sequence in a sample, the method comprising:

• (a) receiving signals from the sample at a relatively high detection temperature and a relatively low detection temperature;
• wherein the first target nucleic acid sequence in the sample is detected by a first signal-generating means and the second target nucleic acid sequence in the sample is detected by a second signal-generating means;
• wherein the detection is performed at one or more cycles of the signal-generating process to obtain a signal value at each of the one or more cycles;
• and wherein the two signals to be generated by the two signal-generating means are not differentiated by a single type of detector;
• (b) processing the signal values obtained in the step (a) by:
• using a second initial reference value to extract the signal for the first target nucleic acid sequence, or using a first initial reference value to extract the signal for the second target nucleic acid sequence;
• wherein the first initial reference value is a value representing a relationship of change in signals provided by the first signal-generating means at the relatively high detection temperature and the relatively low detection temperature, and the second initial reference value is a value representing a relationship of change in signals provided by the second signal-generating means at the relatively high detection temperature and the relatively low detection temperature;
• and
• wherein the first initial reference value is predetermined from a separate reaction using the first target nucleic acid sequence and the first signal-generating means and the second initial reference value is predetermined from a separate reaction using the second target nucleic acid sequence and the second signal-generating means;
• and
• (c) identifying whether the extracted signal satisfies an accuracy criterion;
• when the extracted signal does not satisfy the accuracy criterion, the step (b) is repeated using an amended reference value instead of the initial reference value to re-extract the signal for the first target nucleic acid sequence or the second target nucleic acid sequence;
• wherein the amended reference value is determined such that the re-extracted signal satisfies the accuracy criterion.