Comparative Study of Indirect Fluorescent Antibody, ELISA, and Immunochromatography Tests for Serological Diagnosis of Bovine Babesiosis Caused by Babesia bovis

SIMPLE SUMMARY: Currently serological diagnosis of bovine babesiosis is based on the detection of Babesia-specific antibodies (immunoglobulin-G). Antibody detection is commonly used in seroepidemiological studies or in the assessment of antibabesial antibody titers after cattle vaccination. The indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA) are the most widely used diagnostic tests, although there their implementation has some drawbacks, principally due to the requirements for trained personnel, specific materials, and special laboratory equipment. This study compared a newly designed rapid immunochromatography test (ICT), which has been reported recently and used for Babesia bovis-specific antibody detection with promising results, with an in-house ELISA for the serological diagnosis of cattle exposed to B. bovis (Babesia bovis) in Mexico. Higher sensitivity and specificity values were found by ICT, proving its effectiveness over ELISA. ICT also had better concordance than ELISA when IFAT was used as the “gold standard”. The rapid ICT was shown to have diagnostic utility for the detection of antibodies against B. bovis and could be used as a field test in Mexico due to its practicality, as it does not need laboratory equipment for implementation and interpretation of results. ABSTRACT: The indirect fluorescent antibody test (IFAT) is the most frequently used test to conduct seroepidemiological studies so far, and it is regarded as the "gold standard" test for the serological diagnosis of bovine babesiosis. The aim of the present study was to compare the enzyme-linked immunosorbent assay (ELISA) and the rapid immunochromatography test (ICT) for use in the serological diagnosis of cattle exposed to B. bovis in Mexico. The evaluation of test performance was carried out with 30 positive and 30 negative reference sera. A total of 72 bovine sera samples collected from cattle in a region with endemic bovine babesiosis were analyzed by ELISA and ICT, and the results were compared with those of IFAT. Kappa value (k) was also calculated to determine the agreement between tests. The sensitivity and specificity of ELISA for detecting antibodies against B. bovis were 87% (26/30) and 80% (24/30), respectively. The sensitivity and specificity of ICT for detecting antibodies against B. bovis were 90% (27/30) and 83.3% (25/30), respectively. The overall concordance determined for ELISA and ICT was 94.4% (68/72) and 98.6% (71/72), respectively, when the results were compared with those of IFAT. ICT was more sensitive and specific in this comparative study, showing good strength of agreement (k = 0.79) with respect to IFAT. ICT combines a strip-based assay system that is fast, practical, and sensitive for detection of antibodies to B. bovis, which suggests that it could be applied in the field without requiring any laboratory equipment for its use and interpretation of test results.