Harmful algal blooms in freshwater systems are increasingly common and present threats to drinking water systems, recreational waters, and ecosystems. A highly innovative simple to use, portable biosensor system (MBio) for the rapid and simultaneous detection of multiple cyanobacterial toxins in freshwater is demonstrated. The system utilizes a novel planar waveguide optical sensor that delivers quantitative fluorescent competitive immunoassay results in a disposable cartridge. Data are presented for the world's first duplex microcystin (MC)/cylindrospermopsin (CYN) assay cartridge using a combination of fluorophore-conjugated monoclonal antibodies as detector molecules. The on-cartridge detection limits of 20% inhibitory concentration (IC) was 0.4 μg/L for MC and 0.7 μg/L for CYN. MC assay coverage of eight important MC congeners was demonstrated. Validation using 45 natural lake water samples from Colorado and Lake Erie showed quantitative correlation with commercially available laboratory-based enzyme linked immunosorbent assays. A novel cell lysis module was demonstrated using cyanobacteria cultures. Results show equivalent or better performance than the gold-standard but more tedious 3× freeze-thaw method, with >90% cell lysis for laboratory cultures. The MBio system holds promise as a versatile tool for multiplexed field-based cyanotoxin detection, with future analyte expansion including saxitoxin, anatoxin-a, and marine biotoxins.

Background Point-of-care (POC) CD4 T-cell counting is increasingly recognized as providing improved linkage-to-care during management of HIV infection, particularly in resource-limited settings where disease burden is highest. This study evaluated prototype POC CD4 T-cell counters from MBio Diagnostics in the context of low CD4 count, hospitalized patients in Mozambique. This study measured system performance when presented with challenging, low count samples from HIV/AIDS patients with acute illnesses resulting in hospitalization. Methods Forty whole blood samples were collected from donors on the medical service at Maputo Central Hospital and absolute CD4 counts were generated on the MBio CD4 system and a reference laboratory using flow cytometry. Results The mean and median CD4 counts by the flow cytometry reference were 173 and 80 cells/µL, respectively. Correlation between the MBio CD4 System and the reference was good. Bland-Altman analysis showed a mean bias of +15 cells/µL (+9 to +21 cells/µL, 95% CI), and limits of agreement of −47 to 77 cells/µL. For samples with counts >100 cells/µL (N = 14), the mean coefficient of variation was 7.3%. For samples with counts <50 cells/µL, mean absolute bias of replicate samples was 4.8 cells/µL. When two MBio readers were compared, Bland-Altman bias was −4 cells/µL (−13 to +6 cells/µL, 95% CI), and limits of agreement of −63 and +55 cells/µL. Conclusions The MBio System holds promise as a POC system for quantitation of CD4 T cells in resource-limited settings given system throughput (80–100 cartridges/day), design simplicity, and ease-of-use.

As cyanobacterial harmful algal bloom (cHAB) events increase in scale, severity, frequency, and duration around the world, rapid and accurate monitoring and characterization tools have become critically essential for regulatory and management decision-making. The composition of cHAB-forming cyanobacteria community can change significantly over time and space and be altered by sample preservation and transportation, making in situ monitoring necessary to obtain real-time and localized information. Sandwich hybridization assay (SHA) utilizes capture oligonucleotide probes for sensitive detection of target-specific nucleic acid sequences. As an amplification-free molecular biology technology, SHA can be adapted for in-situ, real-time or near real-time detection and qualitatively or semi-quantitatively monitoring of cHAB-forming cyanobacteria, owing to its characteristics such as being rapid, portable, inexpensive, and amenable to automation, high sensitivity, specificity and robustness, and multiplexing (i.e., detecting multiple targets simultaneously). Despite its successful application in the monitoring of marine and freshwater phytoplankton, there is still room for improvement. The ability to identify a cHAB community rapidly would decrease delays in cyanotoxin analyses, reduce costs, and increase sample throughput, allowing for timely actions to improve environmental and human health and the understanding of short- and long-term bloom dynamics. Real-time detection and quantitation of HAB-forming cyanobacteria is essential for improving environmental and public health and reducing associated costs. We review and propose to apply SHA for in situ cHABs monitoring.