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Microcystins

The Severity of Environmental Contamination Caused by Microcystins

Microcystins (MCs) is a cyclic heptapeptide compound produced by the main cyanobacteria such as Microcystis aeruginosa, Anabaena algae, and Oscillatoria. When cells rupture or age, toxins are released into the water, polluting the environment. MCs is an ecological growth regulator with a self-reinforcing mechanism. High concentrations of MCs will affect the diversity of aquatic plant species, thereby helping cyanobacteria gain a competitive advantage and form water blooms, and the formed water blooms will produce more MCs, Causing a worse impact on the environment. MCs can be detected in 80% of cyanobacteria blooms, and its harm to the water environment and human health has become one of the major environmental issues of global concern.

The Necessity of Monitoring Microcystins with ELISA Testing

Fig. 1 The chemical structure of microcystins

MCs has many isomers, the most common are MC-LR, MC-RR, MC-YR (L, R, Y stand for leucine, arginine, and tyrosine). The most researched at home and abroad are MC-LR and MC-RR. MC has strong stability, heating and boiling can not destroy toxins. Studies have shown that MCs has obvious hepatophilicity, and its pollution is closely related to the occurrence of liver cancer, liver necrosis, and intrahepatic hemorrhage. With continuous in-depth research on the toxicity of MCs, it has also been found that MCs has multiple organ toxicity, genetic toxicity, neurotoxicity, immunotoxicity, and potential cancer-promoting properties, and can cause abnormal development of test organisms. The water polluted by MCs not only has serious toxic effects on humans but also poses a serious threat to environmental sustainability and aquatic life. Therefore, common MCs should be monitored. Due to its high sensitivity, simple sample processing, convenient operation, and lower detection limit than the WHO level, the ELISA detection method is considered to be a more promising method.

ELISA Type

Direct Competitive ELISA

The Advantages of ELISA Testing

Easy to use
Lower cost
High-throughput detection and analysis
High sensitivity and specificity

ELISA Procedure for Microcystins Testing

The microplates were coated with 0.1 mL antibody which was diluted in 0.01M PBS (1:1000 dilution, 10 µg/mL) and incubated at 4°C overnight.

Washed with PBS-Tween (0.35 mL per well; 0.05% Tween 20 in 0.01M PBS), then blocked by BSA-PBS (0.17 mL/well; 0.1% BSA in 0.01M PBS) at 37°C for 30 min.

Washed again with PBS-Tween 4×, followed by incubation with different MCYST analogues (0.05 mL/well in 0.01M PBS) at different dilutions or blank buffer together with the MCYST-HRP conjugate (1:1000 dilution, 25 ng/mL, in 0.01M PBS, 0.05 mL/well) at 37°C for 50 min.

The plate was washed with PBS-Tween 4×, and 0.1 mL of TMB substrate solution was added, incubated at room temperature in the dark for 10 min.

Terminated by adding 0.1 mL of 1 N HCl, the absorbance was determined at 450 nm in the Vmax automatic ELISA reader.

Creative Diagnostics has been committed to algal toxins testing by ELISA. Supported by rich professional knowledge and diversified ELISA kits products, we provide high-quality customized ELISA kits services, professional ELISA testing services, and ELISA development services related to the detection of microcystins. If you wish a lot of careful data, please contact us.

References

Schreidah, CM.; et al. Microcystins: Biogenesis, Toxicity, Analysis, and Control. Chem Res Toxicol. 2020, 33(9): 2225-2246.
Kumar, P.; et al. Recent developments in the methods of quantitative analysis of microcystins. J Biochem Mol Toxicol. 2020, 34(12): e22582.
Yu, FY.; et al. Development of a sensitive ELISA for the determination of microcystins in algae. J Agric Food Chem. 2002, 50(15): 4176-4182.

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