I am performing a (partial) validation of commercially available ELISA kit. I am using the kit on a previously untested matrix, so I wish to validate the assays Accuracy, intra-assay Precision and Spike-recovery.
To my understanding: Accuracy (AC) is defined as: closeness of mean test results to the true concentration. Precision (intra-assay) (PR) is defined as: closeness of agreement (degree of scatter) between a series of measurements obtained from multiple sampling of the same homogenous sample. Spike recovery (SR): used to determine if the assay is affected by the difference between the diluent used to prepare the standard curve and the sample matrix.
Since all these parameters are determined by performing multiple measurements of different concentrations, I am wondering if one can combine these into a single series of samples and use the data to determine the 3 parameters above?
i.e I would perform a series with 4 different concentrations of the analyte spiked into sample matrix, and do 5 determinations for each concentration. Is there any reason that AC, PR and SR determination should be divided into separate series on the validation plate?
I have attached is my planned plate-layout including the other parameters (LOD, LOQ, linearity etc). Am I on the right track here?
I appreciate any thoughts/insight you may have on this