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Evaluation of a magnetic microsphere antibody detection assay for the investigation of exposure to Cryptosporidium parvum and Cryptosporidium hominis
Citation:
Elwin, K., S. Griffin, H. Ward, C. Thornton, R. Puleston, AND R. Chalmers. Evaluation of a magnetic microsphere antibody detection assay for the investigation of exposure to Cryptosporidium parvum and Cryptosporidium hominis. VI International Giardia and Cryptosporidium Conference, Havana, CUBA, April 26 - 27, 2017.
Impact/Purpose:
This poster is for presentation at the VI International Giardia and Cryptosporidium Conference in Havana, Cuba, April 26-27, 2017.
Description:
Anti-Cryptosporidium IgA and IgG are useful markers of exposure to Cryptosporidium in human populations, but detection in saliva may be difficult. To evaluate a magnetic microsphere assay for detection of anti-Cryptosporidium IgA and IgG in saliva, recombinant sporozoite gp15 (15 kDa) proteins from Cryptosporidium hominis and Cryptosporidium parvum coupled to magnetic microspheres were used. Paired serum and saliva samples were collected on multiple occasions in the months following Cryptosporidium diagnosis of ten patients, and the infecting Cryptosporidium species identified in their diagnostic stools. C. hominis and C. parvum gp15 recombinant protein-coupled magnetic beads (BioRad), secondary (detection) goat anti-human IgA and donkey anti-human IgG (Jackson/Stratech), and Streptavidin, R-phycoerythrin conjugate (Invitrogen) were used. The Median Fluorescence Intensity (MFI) of the bead complexes was measured (BioPlex 200 suspension array system, Biorad). Serial dilution of individual patient serum samples indicated that the MFI values decreased at a steady rate in proportion to the dilution of the sample. Variance was low both within and between plates (P>0.05). The saliva responses to the heterogonous recombinant Cryptosporidium species protein were similar to those of the homologous Cryptosporidium species protein (P>0.05), suggesting that either could be used to measure antibody responses to infection with C. hominis or C. parvum. Anti-Cryptosporidium IgG and IgA were detectable in saliva during the six months following diagnosis. A magnetic microsphere antibody assay can be used for the investigation of recent exposure to C. parvum and C. hominis.
