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.
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Department of Pediatrics, Emory University School of Medicine, Atlanta, GA 30322, USA.
Arrowood
M J
MJ
Lejkowski
M
M
Xie
L
L
Schinazi
R F
RF
Mead
J R
JR
D000818:Animals; D002417:Cattle; D002460:Cell Line / Q000469:parasitology; D016785:Cryptosporidium parvum / Q000254:growth & development*; D004285:Dogs; D005069:Evaluation Studies as Topic; D007118:Immunoassay; D007686:Kidney Tubules, Distal / Q000166:cytology; D008163:Luminescent Measurements; D008828:Microbiological Techniques; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity