A chemiluminescence immunoassay for evaluation of Cryptosporidium parvum growth in vitro

A chemiluminescence immunoassay (CLIA) was developed to detect Cryptosporidium parvum growth in Madin-Darby canine kidney (MDCK) cell cultures. Optimal results were obtained when MDCK cells were plated at a density of 1 X 10(4) cells/well (96-well plate) and maintained as a. monolayer for 4 days prior to infection with 2 X 10(4) parasites/well. Two compounds (paromomycin and maduramicin) were evaluated and shown to have selective activity against C. parvum in a dose-dependent manner. There was excellent correlation between CLIA and immunofluorescence assay when assessing anti-C parvum agents in MDCK cells. CLIA offers advantages over conventional enzyme-linked immunosorbent assay and immunofluorescence assay methods in that it is more sensitive and efficient. The combination of,CLIA and MDCK culture provides an efficient tool for evaluating potential anti-cryptosporidial compounds prior to testing in animal models.