Au@Ag Core−Shell Nanoparticles for Colorimetric and Surface- Enhanced Raman-Scattering-Based Multiplex Competitive Lateral Flow Immunoassay for the Simultaneous Detection of Histamine and Parvalbumin in Fish

11 pages, 2 tables, 5 figures.-- This article is licensed under CC-BY 4.0

Foodborne allergies and illnesses represent a major global health concern. In particular, fish can trigger life-threatening food allergic reactions and poisoning effects, mainly caused by the ingestion of parvalbumin toxin. Additionally, preformed histamine in less-than-fresh fish serves as a toxicological alert. Consequently, the analytical assessment of parvalbumin and histamine levels in fish becomes a critical public health safety measure. The multiplex detection of both analytes has emerged as an important issue. The analytical detection of parvalbumin and histamine requires different assays; while the determination of parvalbumin is commonly carried out by enzyme-linked immunosorbent assay, histamine is analyzed by high-performance liquid chromatography. In this study, we present an approach for multiplexing detection and quantification of trace amounts of parvalbumin and histamine in canned fish. This is achieved through a colorimetric and surface-enhanced Raman-scattering-based competitive lateral flow assay (SERS-LFIA) employing plasmonic nanoparticles. Two distinct SERS nanotags tailored for histamine or β-parvalbumin detection were synthesized. Initially, spherical 50 nm Au@Ag core–shell nanoparticles (Au@Ag NPs) were encoded with either rhodamine B isothiocyanate (RBITC) or malachite green isothiocyanate (MGITC). Subsequently, these nanoparticles were bioconjugated with anti-β-parvalbumin and antihistamine, forming the basis for our detection and quantification methodology. Additionally, our approach demonstrates the use of SERS-LFIA for the sensitive and multiplexed detection of parvalbumin and histamine on a single test line, paving the way for on-site detection employing portable Raman instruments

The authors acknowledge financial support from the European Innovation Council (Horizon 2020 Project: 965018-BIOCELLPHE), the MCIN/AEI/10.13039/501100011033 (grant PID2019-108954RB-I00 and PID2019-103845RB-C21), the FSE (“El FSE invierte en tu futuro”), the Xunta de Galicia/FEDER (grant GRC ED431C 2020/09), the European Regional Development Fund (ERDF), and Consejería de Educación y Ciencia del Principado de Asturias (grant ref. SV-PA-21-AYUD/2021/52132). C.F.-L. and L.G.-C. acknowledge Xunta de Galicia for a predoctoral scholarship (Programa de axudas á etapa predoutoral da Consellería de Cultura, Educación e Universidades da Xunta de Galicia, reference number: 2022/294). Funding for open access by the UniversidadedeVigo/CISUG

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