Entity: Universitat Rovira i Virgili (URV)
Confidenciality: No
Education area(s): Nanociència, Materials i Processos - Tecnologia Química de Frontera
Title in different languages: Development of lateral flow assay based on sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of ciguatoxin in fish samples
Abstract: Ciguatera fish poisoning (CFP) is caused by the ingestion of fish contaminated with a marine toxin called ciguatoxin (CTXs). Annually, CFP cases are reported with an estimation of 50,000 and this value will likely increase due to globalization of trade, where contaminated fish disseminated in different areas along with global warming issues will further endanger the public. Avoidance in the consumption of the ciguateric fish is difficult due to the fact that it has the same appearance, smell and taste with the non-‐toxic fish and cannot be inactivated by freezing or cooking. Moreover, currently available methods such as mouse bioassay, analytical and immunoassay methods fall short in addressing reliability, sensitivity and simplicity for detecting CTXs at the point-‐of-‐need at fisheries. Therefore, a reliable, sensitive, and portable assay for routine detection of CTXs is urgently needed. Lateral flow assay (LFA), a commonly implemented assay has gained wide interest in the market due to its simplicity, reliability and accuracy to detect variety of target analytes. Nevertheless, a drawback of this assay can sensitivity in which the format and immunoreagents used plays an important role. In this work, we utilised different nanoparticles such as carbon nanoparticles, gold nanoparticles and latex microspheres as reporter labels and compared their performances. In addition, rabbit IgG was used as a sample target and antibodies against rabbit IgG were utilised as a proof of concept due to the small amounts of CTXs available. Among the particles studied, carbon nanoparticles demonstrated to give the most sensitive results and were then applied to ciguatoxin detection. We successfully developed a LFA based on sandwich ELISA, which can simultaneously detect the four major ciguatoxin congeners (CTX1B, 54-‐deoxyCTX1B, CTX3C and 51-‐hydroxyCTX3C) by mixing the 3G8 and 10C9 monoclonal antibodies as capture probe and the 8H4 antibody conjugated to carbon nanoparticle as a reporter label. The assay was tested using standard samples of CTX1B, and then real samples containing ciguatoxins extracted from fish were applied with a detection range up to 35.66 pg/g of fish sample.
Subject: Enginyeria química
Academic year: 2018-2019
Language: Anglès
Work's public defense date: 2019-09-10
Subject areas: Chemical engineering
Student: Ragasa Reintar, Sharmaine
Work's codirector: Jauset Rubio, Miriam
Department: Enginyeria Química
Creation date in repository: 2021-03-11
TFM credits: 24
Keywords: Ciguatoxin, lateral flow assay, carbon nanoparticles
Title in original language: Development of lateral flow assay based on sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of ciguatoxin in fish samples
Project director: O’Sullivan, Ciara K.
Repositori URV