Application of Au@Pt Nanozyme as Enhancing Label for the Sensitive Lateral Flow Immunoassay of Okadaic Acid.

In this study, a lateral flow immunoassay (LFIA) was developed to detect okadaic acid (OA) belonging to the diarrheic shellfish poisoning group of aquatic toxins. Newly obtained anti-OA monoclonal antibodies and bimetallic core@shell Au@Pt nanoparticles were used in the indirect format of the LFIA. Peroxidase-mimicking nanozyme properties of Au@Pt enabled using them to enhance band coloration on the test strips and, consequently, for increasing the LFIA sensitivity. The instrumental limit of detection (LOD), the working range of detectable concentrations, and the visual cutoff of the assay were 0.5, 0.8-6.8, and 10 ng/mL, respectively. The assay duration was 20 min. The rapid and simple sample preparation procedure was applied for seawater, river water, and fish samples. The total duration of the sample pretreatment and LFIA was 25/40 min for water/fish samples, ensuring testing rapidity. The developed test system provides sensitive control of raw materials and food products and can be used to detect OA at all stages of the food industry «from sea to fork¼ chains.

Triple immunochromatographic test system for detection of priority aquatic toxins in water and fish.

Aquatic toxins are a group of toxic compounds produced by several types of freshwater and marine algae and cyanobacteria and transported through the food chains of water bodies. Potential contamination of aquaculture products (raw and processed fish and seafood) with aquatic toxins requires the use of efficient screening methods for their control. In this study, a multiplex immunochromatographic test system for the simultaneous detection of three aquatic toxins-phycotoxins domoic acid (DA) and okadaic acid (OA), and cyanotoxin microcystin-LR (MC-LR)-is for the first time developed. For this, a competitive indirect immunochromatographic analysis (ICA) based on gold-labeled secondary antibodies was carried out. The LODs/cutoffs/working ranges of the ICA were 0.05/0.3/0.07-0.29, 1.3/100/3.2-58.2, and 0.1/2.0/0.2-1.1 ng/mL for MC-LR, DA, and OA, respectively. The assay duration was 18 min. The developed test system was used to analyze water samples from natural sources (salt and fresh water) and fish samples. For sample preparation of water, simple dilution with a buffer was proposed; for fish samples, methanol-water extraction was utilized. It was demonstrated that the triple LFIA specifically detected target aquatic toxins with recoveries of 85.0-121.5%. The developed multiplex LFIA can be considered a promising analytical solution for the rapid, easy, and sensitive control of water and food safety.

Double Immunochromatographic Test System for Sensitive Detection of Phycotoxins Domoic Acid and Okadaic Acid in Seawater and Seafood.

In this investigation, a double immunochromatographic analysis (ICA) of two relevant phycotoxins, domoic acid (DA) and okadaic acid (OA), was developed for the first time. The ICA was performed in the indirect competitive format using gold nanoparticles conjugated with anti-species antibodies. Under optimal conditions, the instrumental detection limits/cutoffs for simultaneous detection of DA and OA were 1.2/100 and 0.1/2.5 ng/mL, respectively. The time of the assay was 18 min. The ICA was applied to test seawater and a large panel of seafood, including mussels, tiger shrimps, octopuses, whelks, crabs, and scallops. The proposed simple sample preparation method for seafood takes only 20 min. For seawater, a dilution by buffer was implemented. The assay recoveries varied from 80.8% to 124.5%. The competitive potential of the proposed technique as a tool to control natural water and seafood samples is determined by its simplicity, rapidity, and sensitivity.

Rapid detection of phycotoxin domoic acid in seawater and seafood based on the developed lateral flow immunoassay.

A lateral flow immunoassay (LFIA) of phycotoxin domoic acid (DA) contaminating seawater and marine organisms was developed in this investigation. Nine clones of monoclonal antibodies against DA were produced and characterized. The test system was implemented in the indirect competitive format, where gold nanoparticles as a marker were conjugated with secondary antibodies. The developed test system allows for the detection of DA with a cutoff of 60 ng mL-1 and an instrumental detection limit of 1.4 ng mL-1 within 15 min. The LFIA was applied to detect DA in seawater, mussels, shrimps, and octopuses. A simple method of seafood sample preparation was proposed. The entire analytical cycle, from obtaining a sample to the estimation of final results, takes only 30 min. The assay recoveries ranged from 88.5% to 124%. The developed analytical method is a promising solution for rapid on-site monitoring of marine toxicants in water and food throughout the farm-to-fork chain.