A fluorescent immunochromatographic test using immunoliposomes for detecting microcystins and nodularins

Microcystins (MCs), a group of cyclic heptapeptides produced by common cyanobacteria (blue green algae), cause both acute and chronic toxicity. Due to their toxicity, constant monitoring in drinking water, recreational waters as well as other potential exposure through ingestion of contaminated sea food, is very important. In this context, an immunochromatographic test (ICT) using a monoclonal antibody labeled with fluorescent liposomes (immunoliposomes) as tracer was developed, allowing a rapid and simple detection of a large number of MC and nodularin variants in field samples. The present ICT using immunoliposomes proved to be ten times more sensitive than the ICT using colloidal gold for labeling. To achieve quantitative measurement, this ICT was improved by including a stable signal on the control band allowing the expression of the results as a ratio of the fluorescence signals of the specific band versus the control band (SB/CB). Very low concentrations of MC-LR were detected in the analysis buffer (0.06 ng/ml), well below the guideline value of 1 ng/ml proposed by the World Health Organization (WHO), with a dynamic range from 0.06 to 1.5 ng/ml of MC-LR. This method was also validated using a hand-held commercial fluorometer (from ESE (R)), providing the same performances obtained via the analysis station (from Kodak (R)) used in our laboratory. Repeatability tests performed with both devices showed good accuracy (CV<13%). Furthermore, quantification of MCs in natural samples (water bloom and Microcystis culture) was achieved using ICT, leading to similar results obtained via an EIA previously described. All these results demonstrate that this new fluorescent ICT could be used not only as a sensitive detection tool but also to quantify MCs in field samples.