Detection of Cryptosporidium parvum oocysts by dot-blotting using monoclonal antibodies to Cryptosporidium parvum virus 40-kDa capsid protein

J Parasitol. 2008 Feb;94(1):94-8. doi: 10.1645/GE-1313.1.

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. In immunoblotting analysis, MAbCPV40-1 bound to a 40-kDa protein in extracts of C. parvum oocysts. This 40-kDa protein was localized in the sporozoite cytoplasm by immunofluorescence (IFA) staining with MAbCPV40-1. In a dot-blot assay, MAbCPV40-1 was capable of detecting 10(2) non-bleach-treated and 10(2)-10(3) bleach-treated C. parvum oocysts. MAbCPV40-1 was capable of detecting CPV40 antigen in both soluble and total C. parvum oocyst protein extracts, indicating a potential use for detecting this parasite in environmental samples.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology*
  • Capsid Proteins / immunology*
  • Cattle
  • Cryptosporidium parvum / isolation & purification*
  • Cryptosporidium parvum / virology
  • Electrophoresis, Polyacrylamide Gel
  • Fluorescent Antibody Technique
  • Hybridomas
  • Immunoblotting / methods*
  • Mice
  • Mice, Inbred BALB C
  • Oocysts / virology
  • RNA Viruses* / chemistry
  • RNA Viruses* / immunology
  • Recombinant Proteins / immunology
  • Sensitivity and Specificity
  • Symbiosis

Substances

  • Antibodies, Monoclonal
  • Capsid Proteins
  • Recombinant Proteins