Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
867190 | Biosensors and Bioelectronics | 2013 | 8 Pages |
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 °C-treated serum, the limit of detection for this bacterial sensor was lowered by nearly 450-fold, from 31 μg/L to 0.07 μg/L MMC. Real-time quantitative PCR demonstrated that serum-treated cultures responded more strongly to 100 μg/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.
► Bioreporter responses to genotoxins are improved by serum complement activity. ► Mitomycin C can be detected down to 70 parts-per-trillion (ppt). ► This bioreporter system is more sensitive than HPLC for mitomycin C. ► Enhanced effector permeability was verified by fluorescence microscopy/RT-qPCR. ► Demonstrated use within point-of-care diagnostic devices using a centrifugal system.