Abstract
Alkaline phosphatase (ALP) was used as an amplification tool in lateral flow immunoassay (LFIA). Potato virus Х (PVX) was selected as a target analyte because of its high economic importance. Two conjugates of gold nanoparticles were applied, one with mouse monoclonal antibody against PVX and one with ALP-labeled antibody against mouse IgG. They were immobilized to two fiberglass membranes on the test strip for use in LFIA. After exposure to the sample, a substrate for ALP (5-bromo-4-chloro-3-indolyl phosphate/nitro blue tetrazolium) was dropped on the test strip. The insoluble dark-violet diformazan produced by ALP precipitated on the membrane and significantly increased the color intensity of the control and test zones. The limit of detection (0.3 ng mL−1) was 27 times lower than that of conventional LFIA for both buffer and potato leaf extracts. The ALP-enhanced LFIA does not require additional preparation procedures or washing steps and may be used by nontrained persons in resource-limited conditions. The new method of enhancement is highly promising and may lead to application for routine LFIA in different areas.
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The study was financially supported by the Russian Science Foundation (grant 16-16-04108).
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Panferov, V.G., Safenkova, I.V., Varitsev, Y.A. et al. Enhancement of lateral flow immunoassay by alkaline phosphatase: a simple and highly sensitive test for potato virus X. Microchim Acta 185, 25 (2018). https://doi.org/10.1007/s00604-017-2595-3
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DOI: https://doi.org/10.1007/s00604-017-2595-3