The uncertainty and limit of detection in biosensors from immunoassays

This work describes a procedure to obtain the limit of detection (LoD) of a biosensor device from a set of measurements that we call immunoassays, which consists of testing the device's response to different concentrations of the molecule, which has to be detected throughout its working range. The procedure, based on the recommendations of international organizations, provides general expressions to estimate the uncertainty throughout its working range and the LoD. In the metrological model, the contributions to the uncertainty, due to the determination of the parameters of the calibration curve, the resolution, the lack of repeatability and others are taken into account. The model is applied to experimental data from the calibration of a biosensor. Using iterative weighted least squares techniques, a general logistic function (6-parameters) has been fitted and used as a calibration curve. The example studied shows that the lack of repeatability is not always the most important contribution to the LoD. The final expression of the LoD is equivalent to that of the expanded uncertainty (for a coverage probability of 99.9%) assigned to the concentration when . This result permits seeing the LoD as the smallest concentration measured with the device, whereby uncertainty interval does not include negative values. (paper)