Sandwich Hybridization Assay for In Situ Real-Time Cyanobacterial Detection and Monitoring: A Review

Abstract

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.

Tamm Plasmon Polariton Biosensors Based on Porous Silicon: Design, Validation and Analysis

“…The TPP biosensor has the advantages of simplified measurement through reflection spectroscopy without any requirement on polarization selection or wave vector boosting, no need for rinsing of biosensor surface after antigen-antibody reaction, real time monitoring of biomolecular binding process, and CMOS compatible fabrication process. In the future, the sensitivity of the TPP biosensor can be further enhanced by employing sandwich assay which can amplify the level of response significantly [55]. To lower the limit of detection, an advanced spectrometer of 1 pm resolution in wavelength shift can be used.…”

USACE freshwater harmful algal bloom research and development initiative

“…Researchers led by Dr. George Bullerjahn of Bowling Green State University (BGSU) developed a rapid test capable of identifying and quantifying prevalent HAB-causing cyanobacteria in a single water sample at the same time. The multiplexed test employs genera-specific molecular technologies (sandwich hybridization assays [SHAs] targeting 16S rRNA [24] ). To enable on-site use by nonexpert personnel, the molecular technologies are combined with fluorescent/colorimetric probes and field-portable instrumentation.…”

A Digital Twin Lake Framework for Monitoring and Management of Harmful Algal Blooms

“…In addition, the monitoring efficiencies of these methods are quite low and thus they are suitable only for small lakes and reservoirs [17]. To realize efficient monitoring of HABs, many in situ monitoring systems were constructed in lakes [18,19]. With these systems, HAB information and main water quality parameters in core positions were collected continuously and the working costs were reduced.…”