Detection of Viruses By Counting Single Fluorescent Genetically Biotinylated Reporter Immunophage Using a Lateral Flow Assay

Abstract

We demonstrated a lateral flow immunoassay (LFA) for detection of viruses using fluorescently labeled M13 bacteriophage as reporters and single-reporter counting as the readout. AviTag-biotinylated M13 phage were functionalized with antibodies using avidin朾iotin conjugation and fluorescently labeled with AlexaFluor 555. Individual phage bound to target viruses (here MS2 as a model) captured on an LFA membrane strip were imaged using epi-fluorescence microscopy. Using automated image processing, we counted the number of bound phage in micrographs as a function of target concentration. The resultant assay was more sensitive than enzyme-linked immunosorbent assays and traditional colloidal-gold nanoparticle LFAs for direct detection of viruses.

Description

Keywords

bacteriophage, diagnostics, image processing, immunoassay, lateral-flow assay, virus

Citation

Coyright 2015 ACS Appl Mater Interfaces. This is a post-print version of a published paper that is available at: https://pubs.acs.org/doi/abs/10.1021/am5082556. Recommended citation: Kim, Jinsu, Meena Adhikari, Sagar Dhamane, Anna EV Hagstro?m, Katerina Kourentzi, Ulrich Strych, Richard C. Willson, and Jacinta C. Conrad. "Detection of viruses by counting single fluorescent genetically biotinylated reporter immunophage using a lateral flow assay." ACS applied materials & interfaces 7, no. 4 (2015): 2891-2898. DOI: 10.1021/am5082556. This item has been deposited in accordance with publisher copyright and licensing terms and with the author's permission.