title: Detection and Quantification of Toxin-ProducingMicrocystis aeruginosaStrain in Water by NanoGene Assay.
abstract: We demonstrated the quantitative detection of a toxin-producingMicrocystis aeruginosa(M. aeruginosa) strain with the laboratory protocol of the NanoGene assay. The NanoGene assay was selected because its laboratory protocol is in the process of being transplanted into a portable system. ThemcyDgene ofM. aeruginosawas targeted and, as expected, its corresponding fluorescence signal was linearly proportional to themcyDgene copy number. The sensitivity of the NanoGene assay for this purpose was validated using both dsDNAmcyDgene amplicons and genomic DNAs (gDNA). The limit of detection was determined to be 38mcyDgene copies per reaction and 9 algal cells/ml water. The specificity of the assay was also demonstrated by the addition of gDNA extracted from environmental algae into the hybridization reaction. Detection ofM. aeruginosawas performed in the environmental samples with environmentally relevant sensitivity (~105algal cells/ml) and specificity. As expected,M. aeruginosawere not detected in nonspecific environmental algal gDNA over the range of 2 × 100to 2 × 107algal cells/ml.
authors.affiliation: Department of Environmental Science and Engineering, Ewha Womans University, Seoul 03760, Republic of Korea.
authors.forename: Eun-Hee
authors.initials: EH
authors.lastname: Lee
authors.affiliation: Department of Environmental Science and Engineering, Ewha Womans University, Seoul 03760, Republic of Korea.
authors.forename: Kyung-Suk
authors.initials: KS
authors.lastname: Cho
authors.affiliation: Department of Environmental Science and Engineering, Ewha Womans University, Seoul 03760, Republic of Korea.
authors.forename: Ahjeong
authors.initials: A
authors.lastname: Son
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D001427:Bacterial Toxins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D001431:Bacteriological Techniques / Q000295:instrumentation / Q000379:methods*; D004247:DNA / Q000032:analysis; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000302:isolation & purification; D004784:Environmental Monitoring / Q000379:methods; D005798:Genes, Bacterial / Q000235:genetics; D057097:Harmful Algal Bloom; D052998:Microcystins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D046931:Microcystis / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D009693:Nucleic Acid Hybridization / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D014867:Water / Q000737:chemistry; D014871:Water Microbiology
chemical_list: D001426:Bacterial Proteins; D001427:Bacterial Toxins; D017931:DNA Primers; D004269:DNA, Bacterial; D052998:Microcystins; D014867:Water; D004247:DNA
keywords: Algal bloom; Microcystis aeruginosa; NanoGene assay; cyanobacteria; microcystin