碩士 / 國立成功大學 / 環境工程學系碩博士班 / 101 / Cylindrospermopsin (CYN) is a toxic secondary metabolite produced by Cyanobacteria in water body. Because of its stable structure and high solubility in water, CYN is not easy to remove by conventional drinking water treatment processes, resulting in negative effect on human health. CYN has been discovered in a few of Taiwan’s reservoirs, including in main island and in Kinmen island. Therefore, a method to accurately and rapidly estimate the risk associated with the presence of CYN in drinking water sources in required for the protection of public health. Therefore, in this study we proposed to establish an on-site monitoring technology for Cylindrospermopsis cells and CYN-producing cells using qPCR combined with TaqMan system.
Both single and dual quantification systems targeted for the functional genes of Cylindrospermopsis cells, rpoC1, and CYN producing cells, pks, using qPCR combined with TaqMan system were successfully established. The qPCR-TaqMan method was further applied in the on-site monitoring of the targeted genes present in Taiwan’s reservoirs. A total of 73 and 60 samples were collected and analyzed for 11 reservoirs in main island and 11 reservoirs and 2 drinking-water treatment systems in Kingmen island, respectively. During the monitoring, an enzyme-linked immunosorbent assay (ELISA) and an optical microscope were also employed to analyze the concentrations of CYN and Cylindrospermopsis cells, respectively.
Monitoring results showed that good correlations between rpoC1 gene copy numbers detected with the qPCR method and cell numbers enumerated with optical microscope. For another targeted gene, pks, although no correlations were obtained, 90% of the Kinmen samples showed co-occurrence of pks gene copies and CYN concentrations measured with ELISA. A dual quantification system for both pks and rpoC1 genes was also developed. The experimental parameters were tuned to improve the detection of both genes, serving as a basis to better apply the dual detection system in the future.
| Identifer | oai:union.ndltd.org:TW/101NCKU5515161 |
| Date | January 2013 |
| Creators | Ting-HsuanWang, 王婷萱 |
| Contributors | Tsair-Fuh Lin, 林財富 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 114 |
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