title: 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-biotin 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.
authors.affiliation: Chemical and Biomolecular Engineering, ‡Biology and Biochemistry, University of Houston , Houston, Texas 77204, United States.
authors.forename: Jinsu
authors.initials: J
authors.lastname: Kim
authors.forename: Meena
authors.initials: M
authors.lastname: Adhikari
authors.forename: Sagar
authors.initials: S
authors.lastname: Dhamane
authors.forename: Anna E V
authors.initials: AE
authors.lastname: Hagström
authors.forename: Katerina
authors.initials: K
authors.lastname: Kourentzi
authors.forename: Ulrich
authors.initials: U
authors.lastname: Strych
authors.forename: Richard C
authors.initials: RC
authors.lastname: Willson
authors.forename: Jacinta C
authors.initials: JC
authors.lastname: Conrad
mesh_terms: D017104:Bacteriophage M13 / Q000737:chemistry* / Q000302:isolation & purification; D019921:Biotinylation; D005456:Fluorescent Dyes / Q000737:chemistry; D017930:Genes, Reporter; D007118:Immunoassay / Q000295:instrumentation / Q000379:methods*; D013194:Staining and Labeling; D014780:Viruses / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D005456:Fluorescent Dyes
keywords: bacteriophage; diagnostics; image processing; immunoassay; lateral-flow assay; virus