Post-assay growth of gold nanoparticles as a tool for highly sensitive lateral flow immunoassay. Application to the detection of potato virus X.

This article demonstrates a new kind of a highly sensitive lateral flow immunoassay (LFIA). It is based on the enlargement of the size of gold nanoparticles (GNPs) directly on the test strip after a conventional LFIA. Particle size enlargement is accomplished through the catalytic reduction of HAuCl4 in the presence of H2O2 and through the accumulation of additional gold on the surface of the GNPs. To attain maximal enhancement of the coloration of the zone in the test strip and to achieve a minimal background, the concentration of precursors, the pH value, and the incubation time were optimized. GNPs on the test strip are enlarged from 20 to 350 nm after a 1-min treatment at room temperature. The economically important and widespread phytopathogen potato virus X (PVX) was used as the target analyte. The use of the GNP enlargement method results in a 240-fold reduction in the limit of the detection of PVX, which can be as low as 17 pg·mL-1. The total duration of the assay, including virus extraction from the potato leaves, lateral flow, and the enhancement process, is only 12 min. The diagnostic efficiency of the technique was confirmed by its application to the analysis of potato leave samples. No false positives or false negatives were found. The technique does not depend on specific features of the target analyte, and it is conceivably applicable to numerous GNP-based LFIAs for important analytes. Graphical abstract An enlargement solution (containing HAuCl4 and H2O2) was dripped on the strip after common lateral flow immunoassay. Gold nanoparticles on the strip (20 nm) catalyze gold reduction and the formation of larger particles (up to 350 nm), resulting in a 240-fold lower detection limit within 1 min.