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Noncompetitive Chromogenic Lateral-Flow Immunoassay for Simultaneous Detection of Microcystins and Nodularin

Molecular Biotechnology and Diagnostics, Department of Biochemistry, University of Turku, FIN-20520 Turku, Finland
Department of Microbiology, School of Life Sciences, Bharathidasan University, Palkalaiperur, Tiruchirappalli 620024, Tamilnadu, India
Author to whom correspondence should be addressed.
Biosensors 2019, 9(2), 79;
Received: 30 April 2019 / Revised: 4 June 2019 / Accepted: 11 June 2019 / Published: 18 June 2019
(This article belongs to the Special Issue Enzyme-linked Immunoassay)
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Cyanobacterial blooms cause local and global health issues by contaminating surface waters. Microcystins and nodularins are cyclic cyanobacterial peptide toxins comprising numerous natural variants. Most of them are potent hepatotoxins, tumor promoters, and at least microcystin-LR is possibly carcinogenic. In drinking water, the World Health Organization (WHO) recommended the provisional guideline value of 1 µg/L for microcystin-LR. For water used for recreational activity, the guidance values for microcystin concentration varies mostly between 4–25 µg/L in different countries. Current immunoassays or lateral flow strips for microcystin/nodularin are based on indirect competitive method, which are generally more prone to sample interference and sometimes hard to interpret compared to two-site immunoassays. Simple, sensitive, and easy to interpret user-friendly methods for first line screening of microcystin/nodularin near water sources are needed for assessment of water quality and safety. We describe the development of a two-site sandwich format lateral-flow assay for the rapid detection of microcystins and nodularin-R. A unique antibody fragment capable of broadly recognizing immunocomplexes consisting of a capture antibody bound to microcystins/nodularin-R was used to develop the simple lateral flow immunoassay. The assay can visually detect the major hepatotoxins (microcystin-LR, -dmLR, -RR, -dmRR, -YR, -LY, -LF -LW, and nodularin-R) at and below the concentration of 4 µg/L. The signal is directly proportional to the concentration of the respective toxin, and the use of alkaline phosphatase activity offers a cost efficient alternative by eliminating the need of toxin conjugates or other labeling system. The easy to interpret assay has the potential to serve as a microcystins/nodularin screening tool for those involved in water quality monitoring such as municipal authorities, researchers, as well as general public concerned of bathing water quality. View Full-Text
Keywords: noncompetitive immunoassay; cyanotoxin; microcystin; nodularin; lateral flow noncompetitive immunoassay; cyanotoxin; microcystin; nodularin; lateral flow

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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

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Akter, S.; Kustila, T.; Leivo, J.; Muralitharan, G.; Vehniäinen, M.; Lamminmäki, U. Noncompetitive Chromogenic Lateral-Flow Immunoassay for Simultaneous Detection of Microcystins and Nodularin. Biosensors 2019, 9, 79.

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