Comparative Analysis of Machine Learning Algorithms Used for Translating Aptamer-Antigen Binding Kinetic Profiles to Diagnostic Decisions

Current approaches for classifying biosensor data in diagnostics rely on fixed decision thresholds based on receiver operating characteristic (ROC) curves, which can be limited in accuracy for complex and variable signals. To address these limitations, we developed a framework that facilitates the application of machine learning (ML) to diagnostic data for the binary classification of clinical samples, when using real-time electrochemical measurements. The framework was applied to a real-time multimeric aptamer assay (RT-MAp) that captures single-frequency (12.6 Hz) impedance data during the binding of viral protein targets to trimeric aptamers. The impedance data collected from 172 COVID-19 saliva samples were processed through multiple nonlinear regression models to extract nine key features from the transient signals. These features were then used to train three supervised ML algorithms─support vector machine (SVM), artificial neural network (ANN), and random forest (RF)─using a 75:25 training-testing ratio. Traditional ROC-based classification achieved an accuracy of 83.6%, while ML-based models significantly improved performance, with SVM, ANN, and RF achieving accuracies of 86.0%, 100%, and 100%, respectively. The ANN model demonstrated superior performance in handling complex and high-variance biosensor data, providing a robust and scalable solution for improving diagnostic accuracy in point-of-care settings.