Detection of nucleic acids of Cryptosporidium oocyst by surfactant extraction treatment and its application for pathogenic evaluation
Project/Area Number |
25420559
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Research Category |
Grant-in-Aid for Scientific Research (C)
|
Allocation Type | Multi-year Fund |
Section | 一般 |
Research Field |
Civil and environmental engineering
|
Research Institution | University of Shizuoka |
Principal Investigator |
|
Project Period (FY) |
2013-04-01 – 2017-03-31
|
Project Status |
Completed (Fiscal Year 2016)
|
Budget Amount *help |
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2015: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2014: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
|
Keywords | クリプトスポリジウム / 病原性原虫 / リアルタイムPCR / 界面活性剤 / 免疫磁気分離 / 核酸抽出 / オーシスト / 河川水 / 界面活性剤抽出処理 / LAMP法 / 免疫磁気ビーズ / 水道水 / PCR / LAMP / 逆転写酵素 / PCR法 / 飲料水 |
Outline of Final Research Achievements |
Cryptosporidium oocysts are encased in a robust wall that is extremely resistant to environmental stressors such as chlorine used to disinfect potable water. Standard procedures used to extract DNA from oocysts such as the freeze-thaw method with DNA purification kit are time-consuming and expensive. Therefore, we developed the surfactant extraction treatment (SET) that efficiently extracts DNA from the oocyst. Furthermore, we evaluated the ability of SET to detect DNA directly from immunomagnetic bead-oocyst conjugates. As a result, the rate of DNA amplification using immunomagnetic separation-SET (IMS-SET) was greater than that using the standard procedures.
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Report
(5 results)
Research Products
(17 results)