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

doi:10.1021/am5082556

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 journal › Article › peer-review

17 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 language	English (US)
Pages (from-to)	2891-2898
Number of pages	8
Journal	ACS Applied Materials and Interfaces
Volume	7
Issue number	4
DOIs	https://doi.org/10.1021/am5082556
State	Published - Feb 4 2015

Keywords

bacteriophage
diagnostics
image processing
immunoassay
lateral-flow assay
virus

ASJC Scopus subject areas

Materials Science(all)

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10.1021/am5082556

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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.",

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AU - Kim, Jinsu

AU - Adhikari, Meena

AU - Dhamane, Sagar

AU - Hagström, Anna E V

AU - Kourentzi, Katerina

AU - Strych, Ulrich

AU - Willson, Richard C.

AU - Conrad, Jacinta C.

PY - 2015/2/4

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AB - 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.

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KW - image processing

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KW - lateral-flow assay

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

Abstract

Keywords

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