Development of CpGP15 recombinant antigen of Cryptosporidium parvum for detection of the specific antibodies in cattle

The infection of neonatal calves with Cryptosporidium parvum can have a huge economic impact because diarrhea caused by the parasite sometimes results in death. A serodiagnostic system will be helpful in the diagnosis of C. parvum infection. CpP23 is commonly used as an antigen for enzyme-linked immunosorbent assay (ELISA); however, some positive sera show low reactivities, as shown in this study. Herein, we focused on three other antigens, CpGP15, CpP2 and CpGP60, in addition to CpP23, to detect C. parvum-specific antibodies in cattle sera. CpP23 and CpGP15 showed substantial ability to discriminate between positive (n = 10) and negative (n = 10) control cattle sera. Unlike our previous report, both the sensitivity and the specificity were 100% when the two antigens were employed for the ELISA. The newly developed ELISA was applied to a total of 344 sera obtained from 9 cattle farms. Two farms among them had suffered from C. parvum infections before, and were regarded as the C. parvum-positive farms. The positive rates of antibodies against CpP23 and CpGP15 in the C. parvum-positive farms were 42.7% and 49.8%, respectively, whereas the positive rate for either of the antigens was 63.0% in the farms. In contrast, 14.3% and 9.8% were positive for CpP23 and CpGP15 in the C. parvum-negative farms, respectively, whereas 18.8% was positive for either of the antigens. This study revealed that the ELISAs employing both of CpP23 and CpGP15 can avoid false-negative results and are useful for monitoring of the C. parvum infection in cattle farms.