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qPCR assays for the detection and quantification of multiple paralytic shellfish toxin-producing species of Alexandrium

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Ruvindy, R, Bolch, CJ ORCID: 0000-0001-9047-2391, MacKenzie, L, Smith, KF and Murray, SA 2018 , 'qPCR assays for the detection and quantification of multiple paralytic shellfish toxin-producing species of Alexandrium' , Frontiers in Microbiology, vol. 9 , pp. 1-12 , doi: 10.3389/fmicb.2018.03153.

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Abstract

Paralytic shellfish toxin producing dinoflagellates have negatively impacted the shellfish aquaculture industry worldwide, including in Australia and New Zealand. Morphologically identical cryptic species of dinoflagellates that may differ in toxicity, in particular, species of the former Alexandrium tamarense species complex, co-occur in Australia, as they do in multiple regions in Asia and Europe. To understand the dynamics and the ecological drivers of the growth of each species in the field, accurate quantification at the species level is crucial. We have developed the first quantitative polymerase chain reaction (qPCR) primers for A. australiense, and new primers targeting A. ostenfeldii, A. catenella, and A. pacificum. We showed that our new primers for A. pacificum are more specific than previously published primer pairs. These assays can be used to quantify planktonic cells and cysts in the water column and in sediment samples with limits of detection of 2 cells/L for the A. catenella and A. australiense assays, 2 cells/L and 1 cyst/mg sediment for the A. pacificum assay, and 1 cells/L for the A. ostenfeldii assay, and efficiencies of >90%. We utilized these assays to discriminate and quantify co-occurring A. catenella, A. pacificum, and A. australiense in samples from the east coast of Tasmania, Australia.

Item Type: Article
Authors/Creators:Ruvindy, R and Bolch, CJ and MacKenzie, L and Smith, KF and Murray, SA
Keywords: Alexandrium, toxic dinoflagellate, Australia, qPCR detection, qPCR, ribosomal DNA, dinoflagellate cysts
Journal or Publication Title: Frontiers in Microbiology
Publisher: Frontiers Research Foundation
ISSN: 1664-302X
DOI / ID Number: 10.3389/fmicb.2018.03153
Copyright Information:

Copyright 2018 Ruvindy, Bolch, MacKenzie, Smith and Murray. Licensed under Creative Commons Attribution 4.0 International (CC BY 4.0) https://creativecommons.org/licenses/by/4.0/

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