2010 Fiscal Year Final Research Report

Improvement and application of Cryptosporidium detection with Florescent in situ Hybridization

Research Project

Project/Area Number	20760366
Research Category	Grant-in-Aid for Young Scientists (B)
Allocation Type	Single-year Grants
Research Field	Civil and environmental engineering
Research Institution	Anan National College of Technology
Principal Investigator	HASHIMOTO Atsushi Anan National College of Technology, 生命環境学部, 准教授 (30332068)
Project Period (FY)	2008 – 2010
Keywords	Cryptosporidium / FISH
Research Abstract	Fluorescence in situ hybridization (FISH) was applied for specific detection and distinction of Cryptosporidium oocysts by microscopic observation. High fluorescence was observed using the DIAG-CRYR1 probe targeting 18S rRNA partial sequences to distinguish Cryptosporidium spp from Cryptosporidium parvum HNJ-1 oocysts. Combining FISH with fluorescence antibody staining, DAPI, and differential interference contrast observation was valuable for easy differentiation of Cryptosporidium oocysts by microscopic observation. Determination of Cryptosporidium variability by FISH was conducted. Heat inactivated oocysts declined FISH stainability until 100 hours in both PBS and filtrated sewage.

(4 results)