Kim In-Su, Park Hae-Kyung
Nakdong River Environment Research Center, National Institute of Environmental Research, Daegu 43008, Republic of Korea.
Department of Environmental Engineering, Chungbuk National University, Cheongju 28644, Republic of Korea.
Toxins (Basel). 2025 May 12;17(5):242. doi: 10.3390/toxins17050242.
The proliferation of harmful cyanobacteria, particularly , poses significant risks to drinking and recreational water resources, especially under the influence of climate change. Conventional monitoring methods based on microscopy for harmful cyanobacteria management systems are limited in detecting toxigenic genotypes, hindering accurate risk assessment. In this study, we developed a digital droplet PCR (ddPCR)-based method for the simultaneous quantification of total and toxigenic in freshwater environments. We targeted the gene, specific to the genus, and the gene, associated with microcystin biosynthesis. Custom-designed primers and probes showed high specificity and sensitivity, enabling accurate detection without cross-reactivity. The multiplex ddPCR assay allowed for concurrent quantification of both targets in a single reaction, reducing the analysis time and cost. Application to field samples demonstrated good agreement with microscopic counts and revealed seasonal shifts in toxigenic genotype abundance. Notably, ddPCR detected at very low densities-down to 7 cells/mL in the mixed cyanobacterial communities of field samples-even when microscopy failed, highlighting its utility for early bloom detection. This approach provides a reliable and efficient tool for monitoring dynamics and assessing toxin production potential, offering significant advantages for the early warning and proactive management of harmful cyanobacterial blooms.
有害蓝藻的增殖，尤其是[具体蓝藻种类未给出]，对饮用水和娱乐用水资源构成重大风险，特别是在气候变化的影响下。基于显微镜的传统有害蓝藻监测方法在检测产毒基因型方面存在局限性，阻碍了准确的风险评估。在本研究中，我们开发了一种基于数字液滴PCR（ddPCR）的方法，用于同时定量淡水环境中的总[蓝藻种类未给出]和产毒[蓝藻种类未给出]。我们针对特定于[蓝藻属未给出]属的[基因名称未给出]基因以及与微囊藻毒素生物合成相关的[基因名称未给出]基因。定制设计的引物和探针显示出高特异性和灵敏度，能够进行准确检测且无交叉反应。多重ddPCR分析允许在单个反应中同时定量两个目标，减少了分析时间和成本。应用于现场样本显示与显微镜计数结果吻合良好，并揭示了产毒基因型丰度的季节性变化。值得注意的是，即使显微镜检测失败，ddPCR在现场样本的混合蓝藻群落中以非常低的密度（低至7个细胞/毫升）也能检测到[蓝藻种类未给出]，突出了其在早期水华检测中的实用性。这种方法为监测[蓝藻种类未给出]动态和评估毒素产生潜力提供了一种可靠且高效的工具，为有害蓝藻水华的早期预警和主动管理提供了显著优势。
