Development and evaluation of the first immunochromatographic test that can detect specific antibodies against Cryptosporidium parvum

Cryptosporidium parvum is a major cause of diarrhea among human and calves, resulting in severe health hazards and drastic economic losses, respectively. Although C. parvum infection leads to high morbidity and mortality in immunocompromised patients and bovine calves, this infection remains a neglected disease. Currently available diagnostic tests for C. parvum are primarily based on detection of oocysts, DNA, or secreted antigens in fecal specimens. Demonstration of specific antibodies with a rapid immunochromatographic test (ICT) will be advantageous not only in providing a simple, rapid, accurate, and affordable tool but also in surveillance because of the ability to recognize recent and past infections. Herein, we developed two ICTs using the diagnostic antigen CpP23 and immunodominant antigen CpGP15 to detect C. parvum-specific antibodies in cattle sera. Because of unavailability of a reference test for antibody detection, evaluation and validation of our developed ICTs were conducted using reference cattle samples and unknown field cattle sera. Serum samples were simultaneously tested by a previously validated enzyme-linked immunosorbent assay (ELISA) using the same antigens (CpGP15 and CpP23). ICTs showed substantial ability to discriminate between positive and negative control cattle sera for both CpGP15 and CpP23. Even against field sera, high sensitivity, specificity, and agreement rates were recorded for ICTs compared with the previously validated ELISA with the same antigens (CpGP15 = 78.78%, 100%, and 85.11%; CpP23 = 80%, 100%, and 80.56%, respectively). Moreover, a high correlation was observed between the test band intensity of ICTs and optical density of ELISA, particularly in the case of CpP23-specific IgM. To our knowledge, this study represents the first development of ICTs that can detect C. parvum-specific antibodies. Our tests will contribute greatly to C. parvum infection control in cattle by providing a method for on-site diagnosis of early and latent infections.