Abstract:
An intestinal protozoan parasite, Cryptosporidium parvum, is a major cause of waterborne
gastrointestinal disease worldwide. Detection of Cryptosporidium oocysts in potable water is a high
priority for the water treatment industry to reduce potential outbreaks among the consumer
populace. Anti-Cryptosporidium oocyst polyclonal and monoclonal antibodies were tested as
capture and detection reagents for use in a fiber optic biosensor assay for the detection of
Cryptosporidium oocysts. Antibodies were validated using enzyme-linked immunosorbent assays,
flow cytometry, Western blotting and fluorescent microscopy. Oocysts could be detected at a
concentration of 105 oocysts/ml when the polyclonal antibodies were used as the capture and
detection reagents. When oocysts were boiled prior to detection, a ten-fold increase in sensitivity
was achieved using the polyclonal antibody. Western blotting and immunofluorescence revealed
that both the monoclonal and polyclonal antibodies recognize a large (>300 kDa) molecular weight
mucin-like antigen present on the surface of the oocyst wall. The polyclonal antibody also reacted
with a small (105 kDa) molecular weight antigen that was present in boiled samples of oocysts.
Preliminary steps to design an in-line biosensor assay system have shown that oocysts would have
to be concentrated from water samples and heat treated to allow detection by a biosensor assay.
