Comparison of Quantitative Detection Methods for Enumeration
of HAB Species: Applications for Resource Management
There has
been a shift from microscopy to molecular methods
for identifying and quantifying harmful algal bloom (HAB) species in
environmental water samples. Molecular
methods greatly increase the number of samples that can be processed
each day
while also increasing sensitivity and accuracy. While several molecular
methods
have been validated for HAB species quantification, they have not been
rigorously
compared, representing a key gap in our ability to provide
recommendations to
managers for a specific regulatory requirement. Our research provides a
thorough assessment of two molecular methods currently used for HAB
monitoring
and research: quantitative real-time PCR (QPCR) and sandwich
hybridization
assay (SHA).QPCR
and SHA are applicable to several HAB species, but the proposed study
will
focus on the globally-distributed harmful raphidophyte,
Heterosigma
akashiwo
as a model species. Our objectives are to (1) Directly
compare QPCR and SHA
for quantification of
H. akashiwo isolates
spanning a range of cell
abundances, growth phases, and nutrient conditions; (2) Determine the
extent to
which quantification of
H. akashiwo is comparable
using QPCR and SHA for
natural phytoplankton communities (3) Synthesize comparisons according
to a
suite of criteria to enhance HAB monitoring and research activities.
In
this targeted study, QPCR and SHA will be critically compared, and
recommendations
will be made to managers for HAB monitoring strategies based upon
specific
criteria such as the range and limit of detection required, cost per
sample, and
sample throughput. Our results will provide management with the ability
to make
informed decisions when incorporating
molecular
methods into HAB monitoring programs.
Investigators: Dianne Greenfield (dgreenfield@belle.baruch.sc.edu),
University of South Carolina, Belle Baruch Institute for Marine
&
Coastal Sciences and
Kathryn J. Coyne (kcoyne@udel.edu) Funded by a grant from the National
Oceanic and Atmospheric Administration (NOAA).
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