Plasmonic Detection of SARS-CoV-2 Spike Protein with Polymer-Stabilized Glycosylated Gold Nanorods.

The COVID-19 pandemic has highlighted the need for innovative biosensing, diagnostic, and surveillance platforms. Here we report that glycosylated, polymer-stabilized, gold nanorods can bind the SARS-CoV-2 spike protein and show correlation to the presence of SARS-CoV-2 in primary COVID-19 clinical samples. Telechelic polymers were prepared by reversible addition-fragmentation chain-transfer polymerization, enabling the capture of 2,3-sialyllactose and immobilization onto gold nanorods. Control experiments with a panel of lectins and a galactosamine-terminated polymer confirmed the selective binding. The glycosylated rods were shown to give dose-dependent responses against recombinant truncated SARS-CoV-2 spike protein, and the responses were further correlated using primary patient swab samples. The essentiality of the anisotropic particles for reducing the background interference is demonstrated. This highlights the utility of polymer tethering of glycans for plasmonic biosensors of infection.

Glycan-Based Flow-Through Device for the Detection of SARS-COV-2.

The COVID-19 pandemic, and future pandemics, require diagnostic tools to track disease spread and guide the isolation of (a)symptomatic individuals. Lateral-flow diagnostics (LFDs) are rapid and of lower cost than molecular (genetic) tests, with current LFDs using antibodies as their recognition units. Herein, we develop a prototype flow-through device (related, but distinct to LFDs), utilizing N-acetyl neuraminic acid-functionalized, polymer-coated, gold nanoparticles as the detection/capture unit for SARS-COV-2, by targeting the sialic acid-binding site of the spike protein. The prototype device can give rapid results, with higher viral loads being faster than lower viral loads. The prototype's effectiveness is demonstrated using spike protein, lentiviral models, and a panel of heat-inactivated primary patient nasal swabs. The device was also shown to retain detection capability toward recombinant spike proteins from several variants (mutants) of concern. This study provides the proof of principle that glyco-lateral-flow devices could be developed to be used in the tracking monitoring of infectious agents, to complement, or as alternatives to antibody-based systems.

Introduction of a novel COVID-19 biomarker panel by coventry and warwickshire pathology services (CWPS)

COVID-19 is heterogeneous in presentation, with cough, fever, dyspnoea and in some cases, acute respiratory distress syndrome documented. Confidence in the interpretation of clinical symptoms and management of patients can be enhanced with the use of biomarkers and could provide clinicians with a tool to predict prognosis and mortality, allowing for earlier interventions and optimal resource allocation.In March 2020, clinicians approached CWPS requesting the provision of biomarkers, as highlighted in early publications. The aim of this change was to improve the clinical management of patients, remove the need for referral laboratory testing and ensure swift translation of recent evidence into clinical practice. Cost, method availability, IT requirements, assay verification, sample needs and appropriate testing were all considered when extending the scope of service. Continued dialogue with those leading the local COVID clinical pathway ensured the change was clinically supported and that testing was incorporated into the trust ward management strategies.Royal College of Pathology guidelines later published in April 2020 supported the service change and literature reviews continue to highlight the role of inflammatory markers for patient stratification;with a recent systematic review finding some of the included biomarkers increased in more severe infections. The Association of Clinical Biochemistry have also made a statement encouraging this type of innovation, utilising both scientific and medical staff in the improvement of patient care.In collaboration with statisticians from local universities, biomarker data is being interrogated so that any findings may be translated into practice. Currently, multiple regression analysis has allowed the creation of models to explain association of analytes with outcomes and it is hoped continued work will allow the creation of decisions trees and clinical reference values.

Rapid establishment of a COVID-19 biobank at University Hospital Coventry and Warwickshire (UHCW)

Various hypotheses are currently explored regarding COVID-19 and its pathogenesis;however, the clinical spectrum of symptoms, severity and outcomes are not fully understood. Identifying how host response and co-morbidities impact disease presentation and progression are important to enable the development of treatments and predictive markers.In March 2020, Coventry and Warwickshire Pathology Service began saving clinical samples from COVID patients for verification of new assays. Ethical approval was obtained to continue, thus providing a biobank for future collaborative research efforts. Challenges included;the need to establish an effective detection system for samples, the standardisation of procedures to enable timely processing, the organisation of DNA extraction and the storage of samples in an HTA approved facility.Daily search routines were developed to generate lists in a standardised template, enabling staff to identify and retrieve samples quickly. Sample processing was centralised and managed by re-deployed staff. Given supply chain issues with RNA extraction consumables for automated platforms, a manual approach to DNA extraction was taken with the help of local university research staff. Finally, collaboration with the UHCW Arden Tissue Bank enabled the storage of samples, complying with all legislation and regulatory procedures.As a result of the strategies employed, over 10,000 samples have been stored, with numbers continuing to rise. Clinical information has been sourced including;ethnicity, co-morbidities, ventilation, and patient outcome. This has enabled grouping of patients based on disease severity. Since multiple samples from single patients were saved, this has allowed for disease trajectory focussed projects.Not only is the biobank providing samples for trust-led research, through Arden Tissue Bank, samples and ethics can be supplied to academic, commercial and charity organisations - both nationally and internationally.