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Title:A new quantitative PCR assay for detection of potentially anatoxin-producing cyanobacteria
Authors:ID Jablonska, Maša (Author)
ID Eleršek, Tina (Author)
Files:URL URL - Source URL, visit https://doi.org/10.1016/j.hal.2024.102785
 
.pdf PDF - Presentation file, download (1,49 MB)
MD5: 0CF961A0C9F1BFC0059A25F40D86E310
 
Language:English
Typology:1.01 - Original Scientific Article
Organization:Logo NIB - National Institute of Biology
Abstract:Anatoxins (ATX) are globally occurring toxins produced by some species of cyanobacteria in aquatic habitats. They can cause acute poisoning in animals, leading to muscle paralysis and respiratory failure, and might also pose a long-term health risk to humans. Thanks to advances in molecular methods and genomic knowledge, it is now possible to rapidly detect and quantify the genes associated with cyanotoxin production for most major groups of cyanotoxins except ATX. The aim of this study was to develop and validate a new quantitative PCR (qPCR) assay for general detection of all potential ATX producers in the environment. After specificity testing in silico and in vitro with 16 cyanobacterial strains (endpoint PCR, amplicon sequencing and qPCR), two assays targeting the anaC gene were thoroughly validated for linearity, amplification efficiency, sensitivity, dynamic range, inter-assay and intra-assay variability, and the influence of background DNA. The assays were then applied to 144 environmental samples of plankton and biofilm from lakes and rivers whose ATX content had previously been measured by liquid chromatography with tandem mass spectrometry (LC-MS/MS). Amplification efficiency of the two designed assays was between 94% and 103%, and the limits of quantification and detection were up to, but mostly below, 322 and 32 cells/mL, respectively. Both assays showed better or equal specificity in cyanobacterial cultures than currently available PCR assays and were able to predict the presence of ATX detected by LC-MS/MS in most environmental samples (83 % in plankton and 52–62% in biofilm). A higher number of discrepancies between qPCR and LC-MS/MS results in biofilm than in plankton samples indicates limited knowledge and sparse genomic data on benthic cyanobacteria. These assays are the first published general qPCR assays targeting all ATX producers and could provide water managers with a rapid and cost-effective risk assessment to better protect human and animal health.
Keywords:anatoxins (ATX), cyanobacteria, qPCR assay, anaC gene, LC-MS/MS, environmental microbiology, environmental science, toxicology, molecular biology
Publication status:Published
Publication version:Version of Record
Publication date:15.12.2024
Year of publishing:2025
Number of pages:str. 1-12
Numbering:Vol. ǂ142, [art. no.] ǂ102785
PID:20.500.12556/DiRROS-21009 New window
UDC:579
ISSN on article:1878-1470
DOI:10.1016/j.hal.2024.102785 New window
COBISS.SI-ID:219478275 New window
Note:Članek v PDF formatu obsega 55 str.; Online first: 15 Dec. 2024; Nasl. z nasl. zaslona; Opis vira z dne 17. 12. 2024;
Publication date in DiRROS:17.12.2024
Views:259
Downloads:156
Metadata:XML DC-XML DC-RDF
:
JABLONSKA, Maša and ELERŠEK, Tina, 2025, A new quantitative PCR assay for detection of potentially anatoxin-producing cyanobacteria. Harmful algae [online]. 2025. Vol. 142, no. 102785, p. 1–12. [Accessed 21 April 2025]. DOI 10.1016/j.hal.2024.102785. Retrieved from: https://dirros.openscience.si/IzpisGradiva.php?lang=eng&id=21009
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Record is a part of a journal

Title:Harmful algae
Publisher:Elsevier Science
ISSN:1878-1470
COBISS.SI-ID:175350787 New window

Document is financed by a project

Funder:ARIS - Slovenian Research and Innovation Agency
Project number:P1-0245-2019
Name:Ekotoksiologija, toksikološka genomika in karcinogeneza
Funder:ARIS - Slovenian Research and Innovation Agency
Project number:J2-4428-2022
Name:Zelene rešitve za trajnostno večnamensko upravljanje evtrofnih voda
Funder:ARIS - Slovenian Research and Innovation Agency
Funding programme:The Slovenian Research Agency’s Young Researchers Program
Project number:10040146

Licences

License:CC BY 4.0, Creative Commons Attribution 4.0 International
Link:http://creativecommons.org/licenses/by/4.0/
Description:This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.

Secondary language

Language:Slovenian
Keywords:anatoksini (ATX), cianobakterije, qPCR test, gen anaC, okoljska mikrobiologija, znanosti o okolju, toksikologija, molekularna biologija


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