Meiho University Institutional Repository:Item 987654321/1723
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    题名: Rapid on-site multiplex assays for total and toxigenic Microcystis using real-time PCR with microwave cell disruption
    作者: A. Michinaka;H. K. Yen;Y. T. Chiu;H. W. Tsao;T. F. Lin
    贡献者: 健康暨護理學院
    关键词: 16S rDNA;mcyB;microcystins-producing cyanobacteria;multiplex quantitative
    日期: 2012
    上传时间: 2012-09-06T02:29:08Z (UTC)
    摘要: A quantitative real-time polymerase chain reaction (qPCR) is a robust means by which to monitor
    toxin-producing cyanobacteria. However, qPCR usually requires DNA extraction, which is a timeconsuming,
    labor-intensive pretreatment. To be able to quickly determine the potential of cyanotoxin
    contamination in the field, a rapid pretreatment method for DNA extraction and a portable qPCR
    device are needed. In this study, we applied a microwave-based method for the qPCR pretreatment
    and a multicolor portable qPCR with UPL and TaqMan probes to quantify toxigenic and total
    Microcystis. The method was tested using laboratory cultures of toxigenic Microcystis aeruginosa
    PCC7820. The qPCR results showed the cycle thresholds value (Ct value) correlated well with cell
    numbers, with detection limit at about 1,000 cells/ml. This scheme was applied in 22 environmental
    samples from six drinking water reservoirs (DWRs) in Taiwan. Although the results for qPCR were
    about four times higher than those of microscopic observation, good correlation between qPCR and
    microscope methods were obtained (r-square: 0.79, P<0.01). The ratios of toxigenic Microcystis to
    total Microcystis in two reservoirs, Sin-Shan Reservoir and Shih-men Reservoir, were less than 10%.
    In three other reservoirs, Ren-Yi-Tan Reservoir, Nan-Hua Reservoir and Bao-Shan Reservoir, much
    higher (>46.1%) ratios were obtained. The scheme may assist quick assessment of the risk
    associated with toxic cyanobacteria in DWRs.
    關聯: Water Science & Technology
    显示于类别:[生物科技系] 期刊論文

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