Due to the variability of component of assays and analyzers, FibroTest assays can only be performed in validated laboratories [ 91].
INNOTEST® ELISA performance was evaluated in a series of experiments that comprehensively simulated an advanced method validation (i.e., addressing all components of assay performance required by regulatory agencies) (21).
The protein, the main component of the assay, consisted of a fragment of the hemagglutinin of the Influenza A/H1N1 virus (further details are given below).
The Endopep-MS assay relies upon antibody extraction of BoNT/A from a clinical or food sample as a sample preparation step prior to analysis for the toxin, so using the correct antibodies is a critical component of the assay.
Furthermore, the screening results were expected to be affected by interferences arising from factors including compound autofluorescence, noisy/partial concentration responses, false positives acting on the coupled-enzyme component of the assay, and promiscuous inhibitors or potent metal chelators.
A key component of our assay is that survival assessment is irrespective of growth rate as long as they grew into a colony of ∼100 microns after plating.
The rate curve indicated a progressive loss of product formation with complete cessation well before depletion of any component of the assay.
The ability to test expression and molecular mass of the transgenic protein by Western blotting is an essential component of the assay that was facilitated by epitope tagging.
The cleavage product, aminoluciferin (substrate of luciferase), is the component of the assay that generates the luminescent readout that is proportional to caspase-3/7 activity.
Similarly, a primer pair targeted to the gmk gene, which is part of the genotyping component of the assay, also amplifies and distinguishes S. aureus and CoNS.
