Development and Preliminary Evaluation of a Nanoparticle-Assisted PCR Assay for the Detection of Cryptosporidium parvum in Calves

SIMPLE SUMMARY: Accurate and rapid detection of Cryptosporidium parvum is useful for the prevention and control of cryptosporidiosis in humans and animals. This study developed a nano-PCR assay for the rapid detection of C. parvum in calves for the first time, and it was ten-fold more sensitive than the normal PCR assay and had no cross-reaction with other common gastrointestinal pathogens. Further analyses of faecal samples of calves indicated potential usage of the nano-PCR assay in clinical settings. ABSTRACT: C. parvum is an important diarrheal pathogen in humans and animals, especially in young hosts. To accurately and rapidly detect C. parvum infection in calves, we established a nano-PCR assay targeting the cgd3_330 gene for the specific detection of C. parvum. This nano-PCR assay was ten times more sensitive than that of the normal PCR assay by applying the same primers and did not cross-react with C. andersoni, C. bovis, C. ryanae, Balantidium coli, Enterocytozoon bieneusi, Giardia lamblia, and Blastocystis sp. To further test the nano-PCR in clinical settings, a total of 20 faecal samples from calves were examined by using the nano-PCR, the normal PCR, and the nested PCR assays. The positive rates were 30% (6/20), 30% (6/20), and 25% (5/20) for the nano-PCR, the normal PCR, and the nested PCR assays, respectively, indicating that the nano-PCR and the normal PCR assays had the same positive rate (30%). Taken together, the present study could provide a candidate method for the specific detection of C. parvum infection in calves in clinical settings.