Quantitative PCR assay for the simultaneous identification and enumeration of multiple Karenia species

Quantitative PCR (qPCR) is the method of choice for specific detection and quantification of harmful algal bloom (HAB) species. Development of qPCR assay for simultaneous enumeration of species that frequently co-exist in HABs is required. A high sensitivity TaqMan qPCR assay, using probe and primer...

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Bibliographic Details
Published in:Environmental science and pollution research international 2020-10, Vol.27 (29), p.36889-36899
Main Authors: Elleuch, Jihen, Barkallah, Mohamed, Smith, Kirsty F., Ben Neila, Idriss, Fendri, Imen, Abdelkafi, Slim
Format: Article
Language:English
Subjects:
Antimicrobial activity
Antimicrobial agents
Antioxidants
Aquatic Pollution
Atmospheric Protection/Air Quality Control/Air Pollution
Chemical composition
Cineole
Citronellol
Earth and Environmental Science
Ecotoxicology
Environment
Environmental Chemistry
Environmental Health
Environmental science
Essential oils
Gas chromatography
Germacrene
Isoeugenol
Mass spectrometry
Mass spectroscopy
Mollusks
Oils & fats
Ophiolimna mixta
Organic chemistry
Pelargonium
Public health
Research Article
Seafood
Waste Water Technology
Water Management
Water Pollution Control
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Summary:Quantitative PCR (qPCR) is the method of choice for specific detection and quantification of harmful algal bloom (HAB) species. Development of qPCR assay for simultaneous enumeration of species that frequently co-exist in HABs is required. A high sensitivity TaqMan qPCR assay, using probe and primers, located at ITS1–5.8S–ITS2 rDNA region, detecting, specifically, Karenia selliformis , K. bidigitata , and K. mikimotoi , was designed. ITS1–5.8S–ITS2 rDNA region copy numbers per Karenia cell genome were estimated to 217.697 ± 67.904, allowing cell quantification. An application of the designed methodology in field samples has been conducted, and it showed high sensitivity (detection of around 10 −1 cell/100 mg of bivalve mollusk tissue, equivalent to about 20 copies of the target sequence). We suggest that the optimized method could contribute to early detection of three closely related Karenia species in seafood cultivating areas to promote control quality, guarantee a fast and effective intervention, and improve public health prevention.
ISSN:0944-1344
1614-7499
DOI:10.1007/s11356-020-09739-4