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Evaluation of sxtA and rDNA qPCR assays through monitoring of an inshore bloom of Alexandrium catenella Group 1
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Evaluation of sxtA and rDNA qPCR assays through monitoring of an inshore bloom of Alexandrium catenella Group 1
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Journal Article
Evaluation of sxtA and rDNA qPCR assays through monitoring of an inshore bloom of Alexandrium catenella Group 1
2019
Overview
Alexandrium catenella
(formerly
A. tamarense
Group 1, or
A. fundyense
) is the leading cause of Paralytic Shellfish Poisoning in North and South America, Europe, Africa, Australia and Asia. The quantification of
A.catenella
via
sxtA
, a gene involved in Paralytic Shellfish Toxin synthesis, may be a promising approach, but has not been evaluated
in situ
on blooms of
A. catenella
, in which cell abundances may vary from not detectable to in the order of 10
6
cells L
−1
. In this study, we compared
sxtA
assay performance to a qPCR assay targeted to a species-specific region of ribosomal DNA (rDNA) and an established fluorescent
in situ
hybridization (FISH) microscopy method. Passing-Bablok regression analyses revealed the
sxtA
assay to overestimate abundances when <5 cell equivalents
A. catenella
DNA were analysed, but otherwise was closer to microscopy estimates than the rDNA assay, which overestimated abundance across the full range of concentrations analysed, indicative of a copy number difference between the bloom population and a culture used for assay calibration
a priori
. In contrast, the
sxtA
assay performed more consistently, indicating less copy number variation. The
sxtA
assay was generally reliable, fast and effective in quantifying
A. catenella
and was predictive of PST contamination of shellfish.
Publisher
Nature Publishing Group UK,Nature Publishing Group
