Detection of viruses by counting single fluorescent genetically biotinylated reporter immunophage using a lateral flow assay

Jinsu Kim, Meena Adhikari, Sagar Dhamane, Anna E V Hagström, Katerina Kourentzi, Ulrich Strych, Richard C. Willson, Jacinta C. Conrad

Research output: Contribution to journalArticle

15 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)2891-2898
Number of pages8
JournalACS Applied Materials and Interfaces
Volume7
Issue number4
DOIs
StatePublished - Feb 4 2015

Keywords

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

ASJC Scopus subject areas

  • Materials Science(all)

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