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Detection of the Benthic Dinoflagellates, Ostreopsis cf. ovata and Amphidinium massartii (Dinophyceae), Using Loop-Mediated Isothermal Amplification
- Source :
- Journal of Marine Science and Engineering, Volume 9, Issue 8, Journal of Marine Science and Engineering, Vol 9, Iss 885, p 885 (2021)
- Publication Year :
- 2021
- Publisher :
- Multidisciplinary Digital Publishing Institute, 2021.
-
Abstract
- For the in situ and sensitive detection of benthic dinoflagellates, we have established an integrated loop-mediated isothermal amplification (LAMP) assay based on Ostreopsis cf. ovata and Amphidinium massartii. To detect the two species, a set of species-specific primers was constructed between the ITS gene and D1–D6 LSU gene, and the reaction temperature, time, and buffer composition were optimized to establish this method. In addition, the specificity of the LAMP primers was verified both in strains established in the laboratory and in field samples collected from the Jeju coastal waters, Korea. With the LAMP assay, the analysing time was within 45 to 60 min, which may be shorter than that with the conventional PCR. The detection sensitivity of the LAMP assay for O. cf. ovata or A. massartii was comparable to other molecular assays (PCR and quantitative PCR (qPCR)) and microscopy examination. The detection limit of LAMP was 0.1 cell of O. cf. ovata and 1 cell of A. massartii. The optimized LAMP assay was successfully applied to detect O. cf. ovata and A. massartii in field samples. Thus, this study provides an effective method for detecting target benthic dinoflagellate species, and could be further implemented to monitor phytoplankton in field surveys as an altenative.
- Subjects :
- Detection limit
In situ
Chromatography
biology
Amphidinium
Chemistry
Naval architecture. Shipbuilding. Marine engineering
Loop-mediated isothermal amplification
Dinoflagellate
VM1-989
Ocean Engineering
GC1-1581
biology.organism_classification
Oceanography
molecular detection
monitoring
Real-time polymerase chain reaction
Benthic zone
harmful algae
Jeju coastal waters
Water Science and Technology
Civil and Structural Engineering
Dinophyceae
Subjects
Details
- Language :
- English
- ISSN :
- 20771312
- Database :
- OpenAIRE
- Journal :
- Journal of Marine Science and Engineering
- Accession number :
- edsair.doi.dedup.....86ca6b35822053b81bb18ea450d5154e
- Full Text :
- https://doi.org/10.3390/jmse9080885