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Abstract
Four species of Cryptosporidium are routinely found in cattle: Cryptosporidium parvum,
Cryptosporidium bovis, Cryptosporidium ryanae, and Cryptosporidium andersoni. It is
important to determine the species of Cryptosporidium in infected cattle because C.
parvum is the only serious pathogen for humans as well as cattle. Identification of
Cryptosporidium species and genotypes currently relies on molecular methods such as
polymerase chain reaction (PCR) followed by restriction fragment length polymorphism
(RFLP) or gene sequencing. Incorporation of these techniques in a routine veterinary
diagnostic laboratory is cost prohibitive. As such, their applications are limited
primarily to research and a few public health laboratories. To overcome this problem,
a multiplex PCR assay was developed for simultaneously detecting the 4 species of
Cryptosporidium that commonly infect cattle. This assay specifically identifies Cryptosporidium
oocysts present in cattle feces, improves the detection of mixed infections, reduces
the time and cost relative to current sequencing methods, and further demonstrates
the shortcomings of sequencing as the definitive method for identification when analyzing
samples containing mixed infections.