Devices, systems, and methods for high-resolution melt analysis

A method for genotyping a target nucleic acid in a test sample, the method comprising:

• providing a microfluidic device having the test sample and a control sample, the control sample including a wild type of the target nucleic acid;
• providing one or more image-capturing devices configured to acquire images of the test and the control samples to provide high-resolution melt data;
• and
• providing one or more processors coupled to a computer-readable media and in communication with the one or more image-capturing devices, the computer-readable media comprising instructions for:
• obtaining high-resolution melt data from the test sample defining a melting curve for the target nucleic acid in the test sample;
• obtaining high-resolution melt data from the control sample defining a melting curve for the wild type nucleic acid in the control sample;
• calculating melting curve derivatives of the melting curves for the test sample and the control sample, respectively, wherein each melting curve derivative represents a negative derivative of a fluorescence emitted from a nucleic acid sample as a function of temperature causing nucleic acid denaturation;
• calculating parameters defining differences between features of the test sample and the control sample melting curve derivatives;
• and
• assigning a genotype to the test sample based on a comparison of the calculated parameters to predetermined thresholds and boundaries defining genotypes.