The Use of Near-Infrared Light-Emitting Fluorescent Nanodiamond Particles to Detect Ebola Virus Glycoprotein: Technology Development and Proof of Principle

Abstract

Background: There is a dire need for rapid diagnostic tests of high sensitivity, efficiency, and point-of-test reporting capability to mitigate lethal viral epidemic outbreaks.

Purpose: To develop a new operating system within the lateral flow assay (LFA) format for Ebola virus (EBOV), based on fluorescent nanodiamond particles (FNDP) nitrogen vacancy (NV) emitting near-infrared (NIR) light. Specifically, we aimed to detail technical issues and the feasibility of mobilizing FNDP-NV on nitrocellulose membranes (NCM) and capturing them at test and control lines.

Methods: FNDP-NV-200nm, 400nm or 800nm were linked to anti-EBOV glycoprotein (GP) monoclonal antibodies (mAb) and tested for LFA performance by monitoring NIR emissions using an in vivo imaging system or optoelectronic device (OED). Anti-EBOV recombinant glycoprotein (GP) humanized mAb c13C6 was linked to FNDP-NV-200nm for the mobile phase; and a second anti-GP mouse mAb, 6D8, was printed on NCM at the test line. Goat anti-human IgG (GAH-IgG) served as a nonspecific antibody for conjugated FNDP-NV-200nm at the control line.

Results: FNDP-NV-200nm-c13C6 specifically and dose-dependently bound to recombinant EBOV GP in vitro and was effectively captured in a sandwich configuration at the test line by mAb 6D8. FNDP-NV-200nm-c13C6 was captured on the control line by GAH-IgG. The OED quantitative analysis of NIR (obtained in less than 1 minute) was further validated by an in vivo imaging system.

Conclusion: FNDP-NV-200nm performance as a reporter for EBOV GP rapid diagnostic tests suggests an opportunity to replace contemporary visual tests for EBOV GP and other highly lethal viral pathogens. Mobile, battery-operated OED adds portability, quantitative data, rapid data collection, and point-of-test reporting capability. Further development of FNDP-NV-200nm within a LFA format is justified.

Keywords: Ebola virus; LFA; OER; anti-EBOV antibodies; diagnostic lateral flow test; fluidics technology; nitrocellulose membranes; opto-electronic reader.