Next generation planar waveguide detection of microcystins in freshwater and cyanobacterial extracts, utilising a novel lysis method for portable sample preparation and analysis

The study details the development of a fully validated, rapid and portable sensor based method for the on-site analysis of microcystins in freshwater samples. The process employs a novel lysis method for the mechanical lysis of cyanobacterial cells, with glass beads and a handheld frother in only 10 min. The assay utilises an innovative planar waveguide device that, via an evanescent wave excites fluorescent probes, for amplification of signal in a competitive immunoassay, using an anti-microcystin monoclonal with cross-reactivity against the most common, and toxic variants. Validation of the assay showed the limit of detection (LOD) to be 0.78 ng mL−1 and the CCβ to be 1 ng mL−1. Robustness of the assay was demonstrated by intra- and inter-assay testing. Intra-assay analysis had % C.V.s between 8 and 26% and recoveries between 73 and 101%, with inter-assay analysis demonstrating % C.V.s between 5 and 14% and recoveries between 78 and 91%. Comparison with LC–MS/MS showed a high correlation (R2 = 0.9954) between the calculated concentrations of 5 different Microcystis aeruginosa cultures for total microcystin content. Total microcystin content was ascertained by the individual measurement of free and cell-bound microcystins. Free microcystins can be measured to 1 ng mL−1, and with a 10-fold concentration step in the intracellular microcystin protocol (which brings the sample within the range of the calibration curve), intracellular pools may be determined to 0.1 ng mL−1. This allows the determination of microcystins at and below the World Health Organisation (WHO) guideline value of 1 μg L−1. This sensor represents a major advancement in portable analysis capabilities and has the potential for numerous other applications.

Figure optionsDownload as PowerPoint slideHighlights
► Sensitive assay for the detection of the most common and toxic microcystin variants.
► Detection of free and cell bound microcystin for a true reflection of toxin content.
► Novel, highly effective lysis method enabling fast and portable disruption of cells.
► Validated to measure microcystins below 1 and 0.1 ng mL−1; free and intracellular.
► Next generation planar waveguide biosensor combining quantification and ease of use.