RRC ID 27119
Author Hautala H, Lamminmäki U, Spoof L, Nybom S, Meriluoto J, Vehniäinen M.
Title Quantitative PCR detection and improved sample preparation of microcystin-producing Anabaena, Microcystis and Planktothrix.
Journal Ecotoxicol Environ Saf
Abstract Blooms of toxic cyanobacteria, associated with illness and mortality in humans and animals, are becoming increasingly common worldwide. The safe use of surface waters for drinking water production and recreation necessitates assessment of toxigenic cyanobacteria. We have developed simple and reliable sample preparation and qPCR methods to detect microcystin-producing strains of three major bloom-forming genera, Anabaena, Microcystis and Planktothrix. The mcyB second thiolation motif, previously not recognized as a potential target for qPCR, was used as a basis for primer and genus-specific probe design. Assay specificity and sensitivity was confirmed with cultured cyanobacterial strains and the effect of different sample preparation methods on quantification was investigated. Sample filtration and cell lysis reduced assay time and resulted in more efficient amplification compared to DNA extraction. Positive correlation (p<0.005) between mcyB copy numbers and microcystin concentrations was observed in environmental samples. The results encourage the use of qPCR in water risk management.
Volume 87
Pages 49-56
Published 2013-1-1
DOI 10.1016/j.ecoenv.2012.10.008
PII S0147-6513(12)00360-0
PMID 23122919
MeSH Anabaena / classification Anabaena / genetics Animals Cyanobacteria / classification Cyanobacteria / genetics DNA, Bacterial / chemistry Environmental Monitoring / methods* Microcystins / metabolism Microcystis / genetics Real-Time Polymerase Chain Reaction* Risk Management / methods
IF 4.872
Times Cited 19
WOS Category TOXICOLOGY ENVIRONMENTAL SCIENCES
Resource
Algae NIES-107