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Detection and quantification of the toxic microalgae Karenia brevis using lab on a chip mRNA sequence-based
Christos-Moritz Loukas1, Jonathan S McQuillan2, Florian Laouenan3
1National Oceanography Centre, European Way, Southampton SO14 3ZH, United Kingdom; Department of Ocean and Earth Science, University of Southampton Waterfront Campus, European Way, Southampton SO14 3ZH, United Kingdom.
A new Lab Card assay can now detect harmful algae like Karenia brevis in the field. This method uses Nucleic Acid Sequence Based Amplification (NASBA) for sensitive and specific quantification without needing a central lab.
Area of Science:
- Marine Biology
- Biotechnology
- Environmental Science
Background:
- Harmful Algal Blooms (HABs) threaten marine ecosystems and human health.
- Current monitoring methods lack specificity or require centralized laboratory analysis.
- Field-deployable, specific detection of HABs is crucial for timely intervention.
Purpose of the Study:
- To develop and validate a Lab Card-based genetic assay for quantifying the harmful alga Karenia brevis.
- To enable sensitive and specific detection of K. brevis in field conditions.
- To establish a stable, preserved assay for practical field use.
Main Methods:
- Development of a microfluidic Lab Card assay for Karenia brevis quantification.
- Utilized Nucleic Acid Sequence Based Amplification (NASBA) targeting the rbcL gene transcript.
- Combined enzyme dehydration and reagent gelification for preserved Lab Cards with a six-week shelf-life.
Main Results:
- Successfully quantified K. brevis cells (50-5000 cells) using the Lab Card-based rbcL NASBA assay.
- Demonstrated the assay's efficacy with an internal control (IC) for accurate quantification.
- Validated the stability and shelf-life of the preserved Lab Cards.
Conclusions:
- The Lab Card-based IC-NASBA assay provides a sensitive and specific method for quantifying K. brevis in the field.
- This technology overcomes limitations of traditional monitoring, enabling rapid, on-site detection of HABs.
- Represents a significant advancement in HABs monitoring, protecting marine life and human health.
