ORBITEC, in collaboration with Lucigen, proposes a rapid nucleic acid-based detector for spaceflight water systems to enable simultaneous quantification of multiple waterborne pathogens with minimal consumables and crew time. The Rapid Multiplex Microbial Detector (RMMD) amplifies the genetic material in a liquid sample to allow near real-time identification of specific genetic sequences of predetermined bacteria and fungi. This easy-to-use device incorporates a patented polymerase enzyme that enables rapid RNA amplification by reagents with superior long-term shelf life and thermal stability. To operate, a water sample is injected into the RMMD, where it is concentrated and mixed with the reagent. The RMMD is rapidly heated and maintained at an elevated temperature for approximately 15 minutes, then quickly cooled back to room temperature to amplify the genetic material in the sample, which is detected in real time by changes in fluorescence due to dye binding, thus providing quantification. pathogenic cells in the water sample can be rapidly detected quantitatively based on the time of development of fluorescence. Phase 1 activity will result in prototype hardware and software genetic amplification and detection of several pathogenic bacteria and fungi that will bring the technology to TRL 5. The anticipated results of the Phase 2 are an engineering development unit that consists of an amplification/detection process controller, sample cartridges, and reagents, that can be tested in space.