Development and application of an indirect ELISA test for the detection of antibodies to Mycoplasma crocodyli infection in crocodiles (Crocodylus niloticus)

Non-availability of a standardized rapid serodiagnostic test for quick and accurate diagnosis of Mycoplasma crocodyli (M. crocodyli) infection in crocodiles was the underlining reason for conducting the present study. An indirect enzyme-linked immunosorbent assay (iELISA) for the detection of antibodies (Ab) to M. crocodyli infection in crocodile sera was developed using sonicated antigen (Ag) and anti-crocodile conjugate. The iELISA test was optimised with different reagents and at different steps. A cut-off value of percent positive greater than or equal to 53.47% resulted in an estimated sensitivity and specificity of 85.67 and 100%, respectively. The developed iELISA could be used for detection of Abs to M. crocodyli infection in crocodiles and may enable to understand the transmission of the disease.