Objective: To develop a multiplex microbead immunoassay for detection of specific antibodies against Cryptosporidium parvum using recombinant proteins CP23, SA35 and SA40.
Methods: By using purified recombinant proteins CP23, SA35 and SA40 as detected antigens, and bovine serum albumin (BSA) as internal control, the four proteins aforementioned were coupled with micro beads and MIA was developed. Then, the efficiency of the coupled proteins was tested, the difference between the single MIA method and the multiple MIA method was compared, and the difference between plates was also compared.
Results: The purified proteins and BSA were coupled with microbeads successfully, and the MIA method was developed. Finally, the sensitivity and specificity of MIA method were confirmed.
Conclusions: The multiplicate MIA method could be used to detect multiple antibodies after Cryptosporidium parvum infection, and the specificity and sensitivity of MIA are very high. The multiplicate MIA method can be one of the tools used in epidemiological survey.