Simplifying marine biotoxin data analysis: Advancing the Neuro-2a cell-based assay using R

Marine biotoxins (MBs) pose a significant public health risk as seafood contaminants. Testing seafood for MBs before consumption can protect consumers and further provide quality assurances. The in vitro Neuro-2a (N2a) cell-based assay (CBA), is a sensitive, high-throughput, and cost-effective method for the semi-quantification of MBs such as ciguatoxins (CTXs) and brevetoxins (PbTxs), serving as an alternative to animal-based testing. However, the N2a CBA has not been validated, limiting its utility. High data variability based on several potential sources including inter-software inconsistencies, data transfer errors, and improper data processing (calculation errors), or handling (dealing with missing, corrupted, or inconsistent data), have been cited as major challenges to the method. Data errors or inaccuracies can disrupt data integrity and lead to false positive or negative results, with serious health implications. To address these issues, a customized R package, n2a, was developed as the first standardized and free data analysis operating procedure for the N2a CBA. It is designed for simultaneous dose-response analysis of assay derived data and displays parameter estimates, calculates effective concentration (EC₅₀), and generates high-quality graphical outputs. The n2a package ensures rapid dose-response model fitting for larger datasets (2 s per dataset) compared to traditional software (3 min per dataset), increasing data processing efficiency and reducing the potential for data handling errors.