While lateral flow immunoassay (LFIA) is a simple technique that offers a rapid, robust, user friendly, and point-of-care test, its capacity for multiplex detection is rather limited. This study therefore combined the multiplexity of microarray technique and the simple and rapid characteristics of LFIA to enable simultaneous and quantitative detection of five mycotoxins, namely aflatoxin B1 (AFB1), deoxynivalenol (DON), fumonisin B1 (FUMB1), T-2 toxin (T-2), and zearalenone (ZON). In addition, we have synthesized a novel extra-large Stokes shift and strong fluorescence organic compound to be used as a reporter molecule which can be detected under UV light without light filter requirement. Many parameters including microarray spotting buffer, blocking buffer, and concentrations of mycotoxin antibodies were optimized for the microarray LFIA (μLFIA) construction. With the optimal conditions, the μLFIA could accurately and quantitatively detect multiple mycotoxins at the same time. The limits of detection of AFB1, DON, FUMB1, T-2, and ZON were 1.3, 0.5, 0.4, 0.4, and 0.9 ppb, respectively. The recoveries of these five mycotoxins were 70.7%–119.5% and 80.4%–124.8% for intra-assay and inter-assay, respectively. Combining the advantages of the novel reporter molecule and the multiplex capability of μLFIA test, this system could simultaneously detect multiple mycotoxins in one sample with high specificity and high sensitivity. Moreover, this system presents a promising affordable point-of-care platform to detect other analytes as well.
Bibliographical noteFunding Information:
We are grateful to National Center for Genetic Engineering and Biotechnology (Platform Program Grant No. P1550582 and P1950903 ), National Science and Technology Development Agency (Grant No. P1751793 and P1751809 ), and Thailand Science Research and Innovation (Grant No. RSA6280033 ) Thailand, for financial supports of this research.
© 2021 Elsevier B.V.
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- Luminescent organic compound
- Microarray lateral flow strip test
- Multiplex detection
- T-2 toxin
ASJC Scopus subject areas
- Analytical Chemistry