Functionalized viral nanoparticles as ultrasensitive reporters in lateral-flow assays

Meena Adhikari,Anna E V Hagström,Ulrich Strych,Wen-Hsiang Chen,Katerina Kourentzi,Sagar Dhamane,Gavin Garvey,Richard C Willson

doi:10.1039/c3an00891f

Abstract

Two types of viral nanoparticles were functionalized with target-specific antibodies and multiple copies of an enzymatic reporter (horseradish peroxidase). The particles were successfully integrated into an immunochromatographic assay detecting MS2 bacteriophage, a model for viral pathogens. The sensitivity of the assay was greatly superior to conventional gold nanoparticle lateral flow assays, and results could easily be evaluated, even without advanced lab instruments.

Highlights

For MS2 propagation, the E. coli host was propagated in tryptic soy broth (TSB) at 37 C with shaking until log phase
The model analyte used in our study, the E. coli bacteriophage MS2, has been widely used as a model for viral pathogens, and is used as an indicator of water quality by the United States Environmental Protection Agency
Given a sample volume of 100 mL, this corresponds to a titer of 104 pfu mLÀ1, which represents almost one thousand-fold increased sensitivity compared to gold nanoparticle LFA with the same antibodies, and at least a one hundred-fold improvement in sensitivity over previously reported LFAs for viral detection.[4,5]

Summary

Introduction

Two types of viral nanoparticles were functionalized with targetspecific antibodies and multiple copies of an enzymatic reporter (horseradish peroxidase). Functionalized viral nanoparticles have previously been reported to be suitable as affinity reagents in enzyme-linked immunosorbent assays (ELISAs).[6,7,8] Here, we present two different approaches to the manufacture of antibody- and peroxidase-doubly-modi ed M13 bacteriophage, and their use in LFAs (Fig. 1).

Results

Conclusion