Abstract
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.
Highlights
Blooms of cyanobacteria, a phylum of bacteria that obtain their energy through photosynthesis, are commonly found around the globe
The heptapeptide, microcystin in fresh water, and pentapeptide nodularin in brackish water bodies are the potent hepatotoxins acting as specific inhibitors of protein phosphatases (PPs) and hazardous to health, being responsible for growth of tumor precursors, in liver [2,3]
We describe the development of a simple chromogenic lateral-flow immunoassays (LFIA) for simultaneous detection of microcystins and nodularin
Summary
A phylum of bacteria that obtain their energy through photosynthesis, are commonly found around the globe. The most widespread and problematic cyanotoxin group consists of cyclic peptide toxins comprising numerous natural variants. The heptapeptide, microcystin in fresh water, and pentapeptide nodularin in brackish water bodies are the potent hepatotoxins acting as specific inhibitors of protein phosphatases (PPs) and hazardous to health, being responsible for growth of tumor precursors, in liver [2,3]. Microcystin and nodularin are very stable and resistant to hydrolysis, making them a persistent issue for common water usage [5]. The robustness results from the chemical structure of microcystin and nodularin, which consists of a monocyclic peptide ring and a hydrophobic Adda moiety (3-amino-9-methoxy-2,6,8-trimethyl-10-phenyldeca-4(E),6(E)-dienoic acid). The conserved Adda group is the common determinant for all microcystin and nodularin, and has proven useful in the development of broad-specificity detection methods [6,7].
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