Study aimed to estimate presence of Mycobacterium avium subspecies paratuberculosis (MAP) in farm and farmer's goatherds (sacrificed for meat), in Mathura region of North India and to evaluate indigenous ELISA kit, with respect to tissues culture and PCR. Prevalence of Mycobacterium avium subspecies paratuberculosis was 51.7, 37.9, and 46.5% in the goat population in North India, using tissues culture, tissues PCR and ELISA kit, respectively. Prevalence was 65.2, 39.1 and 65.2% in farmers and 42.8, 37.1 and 34.2% in farm herds using culture, PCR and ELISA kit, respectively. In culture MAP was recovered from 39.6 and 32.7% of intestine and MLN tissues, respectively. In PCR 25.8% intestine and 24.1%,MLN were positives. Tissues culture and PCR together detected 62.0% infected goats (51.7% in culture and 37.9% in PCR). In ELISA kit 46.5% goats were positive. Using 3 tests combination (tissues culture and PCR and ELISA kit), 68.9% goats were positive for MAP infection. Whereas, using 2 tests combinations, 62.0, 63.7 and 58.6% goats were positive in Culture and PCR, Culture and ELISA and PCR and ELISA, respectively. In S/P ratio, 46.5% goats were positive in ELISA kit, of which 74.0 and 55.5% were detected in culture and PCR, respectively. Collectively in low positives, positives and strong positives categories, 81.0% goats were detected by kit, of which 53.1 and 44.6% were detected positives in culture and PCR tests, respectively. In farm goats, true prevalence of MAP (culture and PCR) was 54.2% (51.4% goats each were detected in Culture and ELISA and PCR and ELISA combinations). However, in farmer's herds 73.9% goats were positive for MAP infection (true positives). In culture and ELISA and PCR and ELISA, 82.6 and 69.5%, goats were positive, respectively. On applying kappa to direct tissues PCR and ELISA kit with ' tissue culture method, showed 'substantial agreement' both for direct PCR and ELISA kit. ELISA test also showed 'substantial agreement' with direct PCR. Sensitivity and specificity of ELISA with respect to culture and PCR, was 66.6 and 75.0% and 68.1 and 66.6%, respectively. When, PCR was compared with culture, the sensitivity and specificity was 57.1 and 73.3%, respectively. The 39 sheep sacrificed for mutton production were also sampled from slaughterhouse, Mathura. Evaluation of combined data on 97 animals (58 goats and 39 sheep); ELISA kit had 65.4 and 78.5% and 63.4 and 66.0%, sensitivity and specificity with respect to culture and PCR. PCR with culture had 60.0 and 80.9% of sensitivity and specificity, respectively