Fig. 1.

Workflow for analysis of microbiome samples using our paper-based detection platform. Once key bacteria or mRNA targets have been identified, RNA toehold switch sensors and primers for isothermal RNA amplification are designed in silico. Sensors and primers are then rapidly assembled and validated in paper-based reactions. For subsequent use, total RNA is extracted from human fecal samples using a commercially available kit. Specific RNAs are amplified via NASBA (nucleic acid sequence based amplification) and quantified using arrays of toehold switch sensors in paper-based reactions. Microbial and host biomarker RNA concentrations of the samples are determined using a simple calibration curve