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Toward Rapid Detection of Viable Bacteria in Whole Blood for Early Sepsis Diagnostics and Susceptibility Testing
Iyengar, S. N. (Karolinska Institutet (Estocolm, Suècia))
Dietvorst, Jiri (Institut de Microelectrònica de Barcelona)
Ferrer-Vilanova, Amparo (Institut de Microelectrònica de Barcelona)
Guirado López, Gonzalo (Universitat Autònoma de Barcelona. Departament de Química)
Muñoz Berbel, Xavier (Institut de Microelectrònica de Barcelona)
Russom, Aman (Karolinska Institutet)

Data: 2021
Resum: Sepsis is a serious bloodstream infection where the immunity of the host body is compromised, leading to organ failure and death of the patient. In early sepsis, the concentration of bacteria is very low and the time of diagnosis is very critical since mortality increases exponentially with every hour after infection. Common culture-based methods fail in fast bacteria determination, while recent rapid diagnostic methods are expensive and prone to false positives. In this work, we present a sepsis kit for fast detection of bacteria in whole blood, here achieved by combining selective cell lysis and a sensitive colorimetric approach detecting as low as 10 3 CFU/mL bacteria in less than 5 h. Homemade selective cell lysis buffer (combination of saponin and sodium cholate) allows fast processing of whole blood in 5 min while maintaining bacteria alive (100% viability). After filtration, retained bacteria on filter paper are incubated under constant illumination with the electrochromic precursors, i. e. , ferricyanide and ferric ammonium citrate. Viable bacteria metabolically reduce iron(III) complexes, initiating a photocatalytic cascade toward Prussian blue formation. As a proof of concept, we combine this method with antibiotic susceptibility testing to determine the minimum inhibitory concentration (MIC) using two antibiotics (ampicillin and gentamicin). Although this kit is used to demonstrate its applicability to sepsis, this approach is expected to impact other key sectors such as hygiene evaluation, microbial contaminated food/beverage, or UTI, among others.
Ajuts: European Commission 675412
Drets: Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original. Creative Commons
Llengua: Anglès
Document: Article ; recerca ; Versió publicada
Matèria: Sepsis ; Bacteria ; E. coli ; Selective cell lysis ; Prussian blue ; Colorimetric ; Blood
Publicat a: ACS Sensors, Vol. 6 (September 2021) , p. 3357-3366, ISSN 2379-3694

DOI: 10.1021/acssensors.1c01219
PMID: 34410700

10 p, 5.2 MB

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