Hollow nanogold microsphere-signalized lateral flow immunodipstick for the sensitive determination of the neurotoxin brevetoxin B

Abstract

We report on a new membrane-based lateral-flow assay for the rapid determination of the algal biotoxin brevetoxin B (BrevTox) that can be found in fish. Hollow nanogold microspheres (HGMS) with an average size of 45 nm were synthesized by using the reverse micelle method, followed by functionalization with monoclonal mouse anti-BrevTox antibody (mAb-HGMS). They served as the signalling reagent. Bovine serum albumin-BrevTox conjugate and rabbit anti-mouse IgG antibody were manually spotted on the nitrocellulose membrane and served as reagents for the test line and control line, respectively. Both spherical gold nanoparticle-labeled anti-BrevTox antibody (mAb-AuNP) and mAb-HGMS conjugate were investigated with respect to their suitability as signal-transduction tags, and the latter was found to give improved analytical features. The HGMS-based labeling method was then also studied in terms of performance in the immunodipstick assay. Under optimal conditions, the visual detection limit (cut-off value) of the mAb-AuNP based assay is 1.5 ng mL−1, while the sensitivity of the mAb-HGMS based assay is as low as 0.1 ng mL−1. Yes/no decisions can be made within 10 min without the need for expensive instrumentation. The results for the analysis of target BrevTox in spiked fish samples showed a good correlation with data obtained with the commercial ELISA. Importantly, the assay gave no false negative results.