Phage-based assay for rapid detection of bacterial pathogens in blood by Raman spectroscopy

Sepsis is a systemic inflammatory response ensuing from presence and persistence of microorganisms in the bloodstream. The possibility to identify them at low concentrations may improve the problem of human health and therapeutic outcomes. So, sensitive and rapid diagnostic systems are essential to...

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Bibliographic Details
Published in:Journal of immunological methods 2019-02, Vol.465, p.45-52
Main Authors: De Plano, Laura M., Fazio, Enza, Rizzo, Maria Giovanna, Franco, Domenico, Carnazza, Santina, Trusso, Sebastiano, Neri, Fortunato, Guglielmino, Salvatore P.P.
Format: Article
Language:English
Subjects:
Bacteria - classification
Bacteria - growth & development
Bacteria - isolation & purification
Bacteriophage M13 - chemistry
Bacteriophage M13 - immunology
Humans
Immunomagnetic Separation
Limit of Detection
M13 filamentous phage
Peptide Library
Phage display
Phage-capture system
Raman spectroscopy
Sepsis
Sepsis - blood
Sepsis - microbiology
Spectrum Analysis, Raman
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Summary:Sepsis is a systemic inflammatory response ensuing from presence and persistence of microorganisms in the bloodstream. The possibility to identify them at low concentrations may improve the problem of human health and therapeutic outcomes. So, sensitive and rapid diagnostic systems are essential to evaluate bacterial infections during the time, also reducing the cost. In this study, from random M13 phage display libraries, we selected phage clones that specifically bind surface of Staphyloccocus aureus, Pseudomonas aeruginosa and Escherichia coli. Then, commercial magnetic beads were functionalized with phage clones through covalent bond and used as capture and concentrating of pathogens from blood. We found that phage-magnetic beads complex represents a network which enables a cheap, high sensitive and specific detection of the bacteria involved in sepsis by micro-Raman spectroscopy. The enter process required 6 h and has the limit of detection of 10 Colony Forming Units on 7 ml of blood (CFU/7 ml). [Display omitted] •M13 phage clones selected by phage display to specifically bind sepsis pathogens•Phage coated magnetic beads for capture and concentration of few pathogens from blood•Bacteria detection by micro-Raman spectroscopy•Fast, inexpensive and highly sensitive diagnostic system
ISSN:0022-1759
1872-7905
DOI:10.1016/j.jim.2018.12.004