Clostridium difficile (C. difficile) is a prolific pathogen that infects ~500,000 Americans each year and causes severe gastrointestinal stress. C. difficile infection (CDI) often occurs when antibiotics disrupt protective gut microbes, enabling pathogen colonization. CDI is especially problematic in nursing homes where aged residents are frequently overprescribed broad-spectrum antibiotics that can perturb the microbiota. In addition, accurate diagnoses of CDI are challenging as they require outsourced clinical tests. The increased turnaround times for such assays often delay critical treatments and prolong patient suffering. To address these needs, we are developing point-of-care tests for CDI that are readily deployed in nursing homes. These assays comprise engineered, light-emitting enzymes (luciferases) as sensors for secreted proteases from C. difficile. In the absence of the pathogen, the luciferase remains trapped in an inactive conformation, resulting in minimal to no light emission. In the presence of the pathogen, though, the luciferase enzyme is liberated by a C. difficile specific protease and is able to produce photons. The emitted light can be detected by eye or standard cameras, enabling rapid readout of C. difficile presence.

Engineered luciferases as off-the-shelf reporters of pathogenic bacteria

254th National Meeting (2017)

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