A New Statistical Method for Analyzing Elispot Data

The enzyme-linked immunospot assay (ELISpot) is one of the most frequently and widely used methods to detect multiple secretory products from a single cell. There are two main approaches to defining positivity criterion for the quantitative ELISpot assay data: ad hoc (Empirical) rules and hypothesis test methods such as t-test and DFR. Hypothesis tests are indeed important to detect whether the test samples are significantly different from the controls. However, the consistency of the results is also important for clinical tests. This research proposed a new method (called the LOD method) focusing on the reproducibility of the results. Simulation and real data analysis showed that T-test and DFR(eq) have higher test power but LOD method has the highest level of reproducibility.