Abstract: Mycoplasmas cause varied respiratory and genital problems in man and animals. They have great economic importance because of their ability to resist common antibiotics and to survive the cryopreservation of semen during artificial insemination. Mycoplasma equigenitalium is associated with reproductive problems of equines. Though several serological tests can be used for measurement of antibodies to equine-mollicutes, the valuable tests are complement fixation and ELISA. However, ELISA is serotest of choice to detect antibodies in equines against Mycoplasma because of its rapidity, sensitivity and specificity. Capture ELISA could be a useful method for detecting M. equigenitalium antigen in swabs, fluids and tissues where it may not be possible by cultural methods. Here, in the present study, we have done antibody detection by ELISA and antigen detection by capture ELISA. The seroprevalence of M. equigenitalium by indirect ELISA was found to be 20.2% among equines with reproductive problems and only 3.7% in apparently healthy equines. In comparison 4.8% equines with reproductive problems were culturally positive and 9.6% equines were detected positive by capture ELISA. No apparently healthy equid was found positive either culturally or by capture ELISA assay. It shows that indirect antibody ELISA provides a higher possibility of M. equigenitalium antibody detection for mass screening and is less cumbersome than cultural or antigen detection method by capture ELISA.