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Naslov:A new quantitative PCR assay for detection of potentially anatoxin-producing cyanobacteria
Avtorji:ID Jablonska, Maša (Avtor)
ID Eleršek, Tina (Avtor)
Datoteke:URL URL - Izvorni URL, za dostop obiščite https://doi.org/10.1016/j.hal.2024.102785
 
.pdf PDF - Predstavitvena datoteka, prenos (1,49 MB)
MD5: 0CF961A0C9F1BFC0059A25F40D86E310
 
Jezik:Angleški jezik
Tipologija:1.01 - Izvirni znanstveni članek
Organizacija:Logo NIB - Nacionalni inštitut za biologijo
Povzetek: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.
Ključne besede:anatoxins (ATX), cyanobacteria, qPCR assay, anaC gene, LC-MS/MS, environmental microbiology, environmental science, toxicology, molecular biology
Status publikacije:Objavljeno
Verzija publikacije:Objavljena publikacija
Datum objave:15.12.2024
Leto izida:2025
Št. strani:str. 1-12
Številčenje:Vol. ǂ142, [art. no.] ǂ102785
PID:20.500.12556/DiRROS-21009 Novo okno
UDK:579
ISSN pri članku:1878-1470
DOI:10.1016/j.hal.2024.102785 Novo okno
COBISS.SI-ID:219478275 Novo okno
Opomba:Članek v PDF formatu obsega 55 str.; Online first: 15 Dec. 2024; Nasl. z nasl. zaslona; Opis vira z dne 17. 12. 2024;
Datum objave v DiRROS:17.12.2024
Število ogledov:242
Število prenosov:146
Metapodatki:XML DC-XML DC-RDF
:
JABLONSKA, Maša in ELERŠEK, Tina, 2025, A new quantitative PCR assay for detection of potentially anatoxin-producing cyanobacteria. Harmful algae [na spletu]. 2025. Vol. 142, no. 102785, p. 1–12. [Dostopano 8 april 2025]. DOI 10.1016/j.hal.2024.102785. Pridobljeno s: https://dirros.openscience.si/IzpisGradiva.php?lang=slv&id=21009
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Gradivo je del revije

Naslov:Harmful algae
Založnik:Elsevier Science
ISSN:1878-1470
COBISS.SI-ID:175350787 Novo okno

Gradivo je financirano iz projekta

Financer:ARIS - Javna agencija za znanstvenoraziskovalno in inovacijsko dejavnost Republike Slovenije
Številka projekta:P1-0245-2019
Naslov:Ekotoksiologija, toksikološka genomika in karcinogeneza
Financer:ARIS - Javna agencija za znanstvenoraziskovalno in inovacijsko dejavnost Republike Slovenije
Številka projekta:J2-4428-2022
Naslov:Zelene rešitve za trajnostno večnamensko upravljanje evtrofnih voda
Financer:ARIS - Javna agencija za znanstvenoraziskovalno in inovacijsko dejavnost Republike Slovenije
Program financ.:The Slovenian Research Agency’s Young Researchers Program
Številka projekta:10040146

Licence

Licenca:CC BY 4.0, Creative Commons Priznanje avtorstva 4.0 Mednarodna
Povezava:http://creativecommons.org/licenses/by/4.0/deed.sl
Opis:To je standardna licenca Creative Commons, ki daje uporabnikom največ možnosti za nadaljnjo uporabo dela, pri čemer morajo navesti avtorja.

Sekundarni jezik

Jezik:Slovenski jezik
Ključne besede:anatoksini (ATX), cianobakterije, qPCR test, gen anaC, okoljska mikrobiologija, znanosti o okolju, toksikologija, molekularna biologija


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