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Cited by 49Marine Toxins and Detection MethodsMolecular Biology Techniques and ApplicationsMarine Plankton and Biogeochemistry

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Development of a multiplex real-time qPCR assay for simultaneous enumeration of up to four marine toxic bloom-forming microalgal species.

Eckford-Soper, Lisa K., Daugbjerg, Niels·2015·Harmful Algae
DOI 10.1016/j.hal.2015.06.009
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Abstract

Harmful algal blooms (HAB) pose serious economic and health risks worldwide. Current methods of identification require high levels of taxonomic skill and can be highly time-consuming thus limiting sample throughput. So, new rapid and reliable methods for detection and enumeration of HAB species are required. Here we describe a high-throughput, multiplex-qPCR (M-qPCR) method using hydrolysis probe technology for the simultaneous detection of four HAB species commonly found in many coastal areas worldwide: Alexandrium tamarense, Karenia mikimotoi, Karlodinium veneficum and Prymnesium parvum. Primers and probes were species-specific and highly efficient when tested in simplex. Species were then added in succession and the assay conditions adjusted until all four species could be quantitatively evaluated simultaneously. Enumeration accuracy of the M-qPCR assay as a monitoring tool was evaluated using spiked natural environmental samples from Danish coastal waters. Comparison of estimates of cell abundances obtained by the M-qPCR technique with those obtained by light microscopy (Sedgwick Rafter technique) showed no statistically significant difference across a range of concentrations. We were also able to identify and enumerate target cells that would be below the detection limit of light microscopy making this a suitable method for early bloom detection or for low biomass species. With the development of molecular probes for a greater number of algal species M-qPCR will be of great benefit to phytoplankton monitoring programmes and the aquaculture industry worldwide.

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Research works printed but not linked: 28

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Metadata

Authors
Eckford-Soper, L. K., & Daugbjerg, N.
Affiliations
University of Copenhagen (DK)
Corresponding author
Eckford-Soper, Lisa K.
ORCID
Venue
Harmful Algae
Publisher
Elsevier BV
Published
2015-09
Volume
48
Pages
37-43
ISSN
1568-9883 (ISSN-L)
DOI
10.1016/j.hal.2015.06.009
OpenAlex
W990580484
Cited by
49
References
1
Citations by year
2026: 5 · 2025: 3 · 2024: 3 · 2023: 1 · 2022: 8 · 2021: 8 · 2020: 3 · 2019: 9 · 2018: 7 · 2016: 2
FWCI
2.21
Type
journal article
Language
en
Open access
closed
Licence
www.elsevier.com/tdm/userlicense/1.0/ (tdm)
Locations
  • Harmful Algae (publishedVersion)
  • PubMed (publishedVersion)
  • Research at the University of Copenhagen (University of Copenhagen) (submittedVersion)
Funders
Danish Council for Strategic Research (11-116169)
Topics
  • Physical Sciences › Environmental Science › Environmental Chemistry › Marine Toxins and Detection Methods
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Molecular Biology Techniques and Applications
  • Physical Sciences › Earth and Planetary Sciences › Oceanography › Marine Plankton and Biogeochemistry
Keywords
harmful algal blooms, Alexandrium tamarense, Karenia mikimotoi, Karlodinium veneficum, Prymnesium parvum, TaqMan probes, phytoplankton monitoring, aquaculture, coastal waters, light microscopy comparison

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Eckford-Soper, L. K., & Daugbjerg, N. (2015). Development of a multiplex real-time qPCR assay for simultaneous enumeration of up to four marine toxic bloom-forming microalgal species. Harmful Algae. https://doi.org/10.1016/j.hal.2015.06.009