Multiple simultaneous detection of Harmful Algal Blooms (HABs) through a high throughput bead array technology, with potential use in phytoplankton community analysis

G. Scorzetti, Larry E Brand, G. L. Hitchcock, K. S. Rein, C. D. Sinigalliano, J. W. Fell

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

As an alternative to traditional, morphology-based methods, molecular techniques can provide detection of multiple species within the HAB community and, more widely, the phytoplankton community in a rapid, accurate and simultaneous qualitative analysis. These methods require detailed knowledge of the molecular diversity within taxa in order to design efficient specific primers and specific probes able to avoid cross-reaction with non-target sequences. Isolates from Florida coastal communities were sequence-analyzed and compared with the GenBank database. Almost 44% of the genotypes obtained did not match any sequence in GenBank, showing the existence of a large and still unexplored biodiversity among taxa. Based on these results and on the GenBank database, we designed 14 species-specific probes and 4 sets of specific primers. Multiple simultaneous detection was achieved with a bead array method based on the use of a flow cytometer and color-coded microspheres, which are conjugated to the developed probes. Following a parallel double PCR amplification, which employed universal primers in a singleplex reaction and a set of species-specific primers in multiplex, detection was performed in a cost effective and highly specific analysis. This multi-format assay, which required less than 4 h to complete from sample collection, can be expanded according to need. Up to 100 different species can be identified simultaneously in a single sample, which allows for additional use of this method in community analyses extended to all phytoplankton species. Our initial field trials, which were based on the 14 species-specific probes, showed the co-existence and dominance of two or more species of Karenia during toxic blooms in Florida waters.

Original languageEnglish
Pages (from-to)196-211
Number of pages16
JournalHarmful Algae
Volume8
Issue number2
DOIs
StatePublished - Jan 1 2009

Fingerprint

algal blooms
algal bloom
phytoplankton
probes (equipment)
probe
methodology
qualitative analysis
cross reaction
field experimentation
detection
analysis
biodiversity
coexistence
sampling
amplification
color
genotype
assays
assay
method

Keywords

  • Bead array
  • Harmful Algal Bloom
  • Luminex
  • Simultaneous molecular detection

ASJC Scopus subject areas

  • Aquatic Science
  • Plant Science

Cite this

Multiple simultaneous detection of Harmful Algal Blooms (HABs) through a high throughput bead array technology, with potential use in phytoplankton community analysis. / Scorzetti, G.; Brand, Larry E; Hitchcock, G. L.; Rein, K. S.; Sinigalliano, C. D.; Fell, J. W.

In: Harmful Algae, Vol. 8, No. 2, 01.01.2009, p. 196-211.

Research output: Contribution to journalArticle

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