Smartphone-based pathogen diagnosis in urinary sepsis patients

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Smartphone-based pathogen diagnosis in urinary sepsis patients Lucien Barnes, Douglas M. Heithoff, Scott P. Mahan, Gary N. Fox, Andrea Zambrano, Jane Choe, Lynn N. Fitzgibbons, Jamey D. Marth, Jeffrey C. Fried, H. Tom Soh, Michael J. Mahan EBioMedicine Volume 36, Pages 73-82 (October 2018) DOI: 10.1016/j.ebiom.2018.09.001 Copyright © 2018 The Authors Terms and Conditions

EBioMedicine 2018 36, 73-82DOI: (10.1016/j.ebiom.2018.09.001) Copyright © 2018 The Authors Terms and Conditions

Fig. 1 SmaRT-LAMP direct specimen testing of urine from sepsis patients. (a) Assay schematic for smaRT-LAMP, which entails sample collection, bacterial cell lysis/reagent addition, and real-time analysis via the smartphone. (b) Schematic of workflow for the Bacticount app, which analyzes fluorescence data collected continuously from multiple samples through the phone's camera (left panel), and then uses these data to automatically determine the genome copy-number of bacterial pathogens in real time (right panel). EBioMedicine 2018 36, 73-82DOI: (10.1016/j.ebiom.2018.09.001) Copyright © 2018 The Authors Terms and Conditions

Fig. 2 SmaRT-LAMP quantification of ST with performance equivalent to a benchtop laboratory qPCR instrument. (a–d) Normalized representative traces and Tt values of ST CFU in buffer at concentrations of 101−105 CFU/mL using qPCR-LAMP and smaRT-LAMP; (e, f), Percentage of total samples amplified at each concentration using qPCR-LAMP or smaRT-LAMP (21 samples/concentration). EBioMedicine 2018 36, 73-82DOI: (10.1016/j.ebiom.2018.09.001) Copyright © 2018 The Authors Terms and Conditions

Fig. 3 SmaRT-LAMP quantification of ST in spiked diverse biological specimens. (a–f) Tt values for ST CFU in murine blood, urine, and feces using qPCR-LAMP and smaRT-LAMP. (g, h) corresponding representative traces (2–2 × 104 CFU/reaction); NC, no cell. (I, J) Percentage of total pathogen samples amplifying at each concentration using smaRT-LAMP. EBioMedicine 2018 36, 73-82DOI: (10.1016/j.ebiom.2018.09.001) Copyright © 2018 The Authors Terms and Conditions

Fig. 4 SmaRT-LAMP quantitation of diverse pathogens in spiked murine whole blood and human donor urine. (a–e) Tt values for SPN, SE, EC, YP (2–2 × 104 CFU/reaction); SA (4 × 101–2 × 104 CFU/reaction). (f) Percentage of total pathogen samples amplifying at each concentration in smaRT-LAMP. (g, h) Representative traces and Tt values for EC in spiked human donor urine (2–2 × 104 CFU/reaction). (i) Percentage of total EC samples amplifying at each concentration in smaRT-LAMP (≥ 10 samples/concentration). EBioMedicine 2018 36, 73-82DOI: (10.1016/j.ebiom.2018.09.001) Copyright © 2018 The Authors Terms and Conditions

Fig. 5 SmaRT-LAMP detection and quantitation of Salmonella in whole blood of septic mice. (a–c). Mice were orally infected with ST via gastric intubation at a dose of 2 × 107 cells and whole blood was sampled at days 6 (pre-sepsis), 8 (sepsis), and 10 (severe sepsis) post-infection. CFU were determined by qPCR-LAMP (closed boxes), smaRT-LAMP (open boxes), and direct colony count (circles). n = 14 mice. EBioMedicine 2018 36, 73-82DOI: (10.1016/j.ebiom.2018.09.001) Copyright © 2018 The Authors Terms and Conditions