Whichever path they choose, assay developers must address several problems to produce useful toxicology tools.
We sought to assess a new point-of-care assay the Dual Path Platform (DPP) syphilis assay, which is based on simultaneous detection of antibodies to treponemal and non-treponemal antigens for guiding use of antibiotics for yaws eradication.
Recently, other serologic tests have been developed – MAPIA, RT and dual-path-platform (DPP) VetTB assay – using M. bovis-specific antigens such as MPB83, ESAT6 and CFP-10 antigens [ 18, 26, 41].
In the PathHunter® U2OS Keap1–Nrfunctionalnassaysay that uses β-galactosidase-based enzyme fragment complementation technology and luminescence for the detection of Nrf2 nuclear translocation, LH601A promoted the nuclear translocation of Nrf2 with a similar EC50 of 12 μmol/L.
This compares favorably with other embodiments of absorbance detection in microfluidic systems (e.g., 13 μM with a 28 μm path length) and, for the assay presented herein, is more than sufficient to extract quantitative information (from enzyme turnovers that give rise to product concentrations in the micromolar to millimolar range).
The fluorescein was used as a passive reference to correct for light path anomalies in the microplate assay and detected by exciting at 480 nm and reading the emission at 520 nm; the bis-ANS measurement was normalized against this value.
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While the latter certainly is a challenge for rare diseases, we have yet recently shown a path of successfully translating 'discovery' assays into clinically more applicable tests (Qundos et al, 2014).
Recent studies have demonstrated the advantages of using multiple antigens (e.g. ESAT-6, CFP10 and MPB83) in multi-antigen print immune-assay (MAPIA) [ 12, 26], lateral flow rapid test (RT) [ 18] or dual path platform (DPP) [ 27] assays.
Whereas in the past assays were predominantly in vitro using purified components, there is now a realization that cell-based, high-content assays are a more productive path.
