Development of a method for quantification of Cyclospora cayetanensis in environmental water

Research Project

Project/Area Number	25630223
Research Category	Grant-in-Aid for Challenging Exploratory Research
Allocation Type	Multi-year Fund
Research Field	Civil and environmental engineering
Research Institution	University of Yamanashi
Principal Investigator	HARAMOTO Eiji 山梨大学, 総合研究部, 准教授 (00401141)
Research Collaborator	KITAJIMA Masaaki Singapore-MIT Alliance for Research and Technology
Project Period (FY)	2013-04-01 – 2015-03-31
Project Status	Completed (Fiscal Year 2014)
Budget Amount *help	¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000) Fiscal Year 2014: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000) Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywords	健康関連微生物 / サイクロスポーラ / 原虫
Outline of Final Research Achievements	This study aimed to develop a highly specific and sensitive quantitative PCR (qPCR) assay for Cyclospora cayetanensis genomes. SYBR Green-based qPCR assay targeting ITS-2 gene of C. cayetanensis could detect quantitatively as low as 100 copies/reaction of artificially synthesized plasmid DNA containing the target sequence for C. cayetanensis. The developed qPCR assay was successfully used to detect indigenous C. cayetanensis genomes in 6 (25%) of 24 influent and 3 (13%) of 24 effluent samples collected monthly during a year at two wastewater treatment plants in Arizona, U.S.A., with the highest concentration of 12000 copies/L.