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Title: Detection of Cryptosporidium parvum oocysts by dot-blotting using monoclonal antibodies to CPV40 capsid protein

item Jenkins, Mark
item Obrien, Celia
item Trout, James

Submitted to: Journal of Parasitology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/17/2007
Publication Date: 2/5/2008
Citation: Jenkins, M.C., Obrien, C.N., Trout, J.M. 2008. Detection of Cryptosporidium parvum oocysts by dot-blotting using monoclonal antibodies to CPV40 capsid protein. Journal of Parasitology. 94:94-98.

Interpretive Summary: Cryptosporidiosis is an intestinal parasitic disease of humans and calves caused by species of Cryptosporidium. The parasite is spread through a fecal-oral route of infection. Preventing outbreaks of cryptosporidiosis depends in part on rapidly identifying the parasite in water that is destined for consumption. Although a number of tests exist for identifying Cryptosporidium oocysts, these methods are generally insensitive to levels of the parasite that can cause infection. The present study describes a monoclonal antibody (MAb) that recognizes a virus inside C. parvum oocysts. This MAb is capable of detecting the organism in water samples using a dot-blot assay. A test based on this MAb maybe useful for rapid and sensitive detection of C. parvum oocysts in water.

Technical Abstract: Monoclonal antibodies (MAb) were prepared against the 40 kDa capsid protein of Cryptosporidium parvum virus (CPV) by immunizing mice with purified recombinant CPV40 protein. By immunoblotting analysis, one MAb, designated MAbCPV40-1, bound to a 40 kDa protein in extracts of C. parvum oocysts, which in immunofluorscence antibody staining localized in the sporozoite cytoplasm. In a dot blot assay, MAbCPV40-1 was capable of detecting between 101 and 102 purified C. parvum oocysts. Surface water (10 l) that had been spiked with C. parvum oocysts before or after filtration was processed for total particulates using Envirochek columns in a standard concentration method. By dot blot assay, MAbCPV40-1 detected 102 - 103 recovered C. parvum oocysts indicating the potential use of this MAb reagent for detecting C. parvum in surface waters.