Electrochemical biosensor development for enhanced detection of pathogens which cause sepsis and hyperinflammation

Vezza, Vincent

Thesis

Electrochemical biosensor development for enhanced detection of pathogens which cause sepsis and hyperinflammation

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Creator

Vezza, Vincent

Rights statement

Strathclyde Thesis Copyright

Awarding institution

University of Strathclyde

Date of award

2021

Thesis identifier

T16067

Person Identifier (Local)

201677757

Qualification Level

Doctoral (Postgraduate)

Qualification Name

Doctor of Engineering (EngD)

Department, School or Faculty

Department of Biomedical Engineering

Abstract

Sepsis represents a significant and growing challenge within modern healthcare settings. It can deteriorate quickly, potentially leading to fatality. The common characteristic to all sepsis patients is the importance of pathogen identification. Currently, pathogen detection is lengthy (hours to days) and as such is detrimental to patient outcome. Rapid pathogen detection will aid clinicians in source control and help guide targeted treatment in shorter time scales giving patients the best chances for recovery. Electrochemical biosensors have potential to offer a route to rapid, low cost, multiplexed pathogen identification. The work presented in this thesis represents a series of investigations made to apply electrochemical methods to rapid pathogen detection at the point of care. The first section presents a literature review exploring sepsis, current detection methods employed and identifying the requirements of an electrochemical biosensor for pathogen detection. The second section details investigations into suitable platforms on which to construct electrochemical biosensors. Various electrode platforms were investigated and the results from these experiments were used to design custom ‘printed circuit board’ electrodes. The ability to form sensing layers composed of biorecognition elements on printed circuit board electrode surfaces is then demonstrated using Streptococcus pneumoniae genomic DNA to form a S. pneumoniae recognition surface. lytA was detectable on this S. pneumoniae DNA biosensor in serum samples at 1 pM after 15 minutes of sample addition at room temperature. Finally, the development of a simple enzyme-based sensor for SARS-CoV-2 is demonstrated. Detection performances are described for multiple positive and negative controls culminating in clinical sample testing. Viral PCR sample solution clinical samples are shown to be statistically different from a negative sample (P = 1.2E-7). Viral transport medium clinical samples showed multiple positive and negative samples were significantly different with preliminary calculations showing a sensitivity = 80% and specificity = 67%.

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