Field portable Coliform bacteria & E. Coli RNA biomarker LAMP - OSD system

Fabrico Technology, Dr. Sanchita Bhadra and Professor Andrew Ellington of the University of Texas at Austin, have successfully demonstrated the proof-of-concept of an unparalleled, first-in-class in-field nucleic acid diagnostic platform for distinction of viable from non-viable E. coli and coliforms in <5 h. This platform is based on a novel variation of the Ellington patent-pending loop-mediated isothermal amplification (LAMP) - oligonucleotide strand displacing (OSD) assay, which is capable of not only exquisitely sequence-specific analyte detection but also analyte copy number estimation with only single endpoint visual 'yes/no' readout of presence or absence of signal. By comparing ratiometrically the increase in nucleic acid analyte copies in live bacteria and lack thereof in dead counterparts our thresholded RT-LAMP-OSD System allows for highly specific 'Yes/No' answers with minimal processing time, and technical and infrastructure complexities.The Phase I demonstrated that the thresholded RT-LAMP-OSD assay can meet the detection limit of 1 CFU/100 mL and is highly specific to only coliforms and E. coli.In Phase II, we propose to optimize and validate specificity of our thresholded RT-LAMP-OSD System by testing against background bacteria, test for chlorine-stressed E. coli and coliform, and prepare an EPA ATP application for independent laboratory verification of the system.