Serum complement enhances the responses of genotoxin- and oxidative stress-sensitive Escherichia coli bioreporters

Abstract

Bacterial bioreporters are limited in their abilities to detect large polar molecules due to their membrane selectivity. In this study, the activity of serum complement was used to bypass this undesired selectivity. Initially, the serum complement activity was assessed using the responses of a bacterial bioreporter harboring a recA::luxCDABE transcriptional fusion when exposed to the chemotherapy drug, mitomycin C (MMC). Using 50 degrees C-treated serum, the limit of detection for this bacterial sensor was lowered by nearly 450-fold, from 31 mu g/L to 0.07 mu g/L MMC. Real-time quantitative PCR demonstrated that serum-treated cultures responded more strongly to 100 mu g/L MMC, with 3.1-fold higher recA expression levels. Subsequent experiments with other bioreporter strains also found enhanced sensitivities and responses. Finally, combining each of the above findings, tests were performed to demonstrate the potential application of the recA::luxCDABE bioreporter within a lab-on-a-CD platform as a point-of-care diagnostic to measure chemotherapeutic drug concentrations within blood.