Three ELISA kits were compared for screening of animals and human beings in India. Kit 1: Indigenous ELISA kit had protoplasmic antigen (PA) from MAP 'Bison type' of goat origin. Kit 2: Antigen (PA) of Kit 1 was replaced with commercial purified protoplasmic antigen (PPA) of MAP 'bovine' origin (Allied Monitor Inc., USA). Kit 3: Commercial ELISA kit for bovines (Pouquier, France). Kit 1 was used as. Serum ELISA kit 1 (s-Kit 1) and milk-ELISA kit 1 (m-Kit 1). Overall sensitivity and specificity of s-Kit 1 with culture (Feces/milk/tissues) was between 28.5-95.6% and 50.0-90.6%, respectively. M-Kit 1 with respect to culture (milk and feces), sensitivity and specificity ranged between 28.5-91.9% and 50.0-75.0%, respectively. Using s-Kit 1 in homologous host, the sensitivity and specificity was 55.5 and 86.3% with respect to fecal culture and were 66.6 and 75.0% with respect to tissues culture, respectively. In advance stages of Johne's disease in a private farm (35 goats and kids with 100% morbidity due JD) with respect to fecal culture, the sensitivity and specificity was, 37.0 and 50.0%, respectively. However, in sheep, the sensitivity and specificity of s-Kit 1 with respect to fecal and tissues culture was variable between 40.0-68.7% and 75.0-90.0%, respectively. In cattle, s-Kit 1, had 50.0 and 90.6%, sensitivity and specificity with respect to fecal culture With respect to milk culture it had sensitivity of 95.6% and specificity could not be determined due to lack of negative samples. Using m-Kit 1, in goats, sensitivity and specificity was, 56.7 and 50.0%, respectively, with respect to milk culture. In cattle, with respect to milk culture, the sensitivity and specificity was 28.5 and 75.0%, respectively. Low correlation with ELISA and culture was due to low conversion of serum globulins to lacto-globulins. In another studies, sensitivities were 90.0 and 90.9%, respectively in milk and fecal culture. However, in human beings, using s-Kit 1, sensitivity and specificity were, 100.0 and 33.3%, respectively. PA in s-Kit 1, detected 10.5 and 46.7% kids positives in farmer's herds, whereas Kit 2 detected, nil and 2.7% kids, respectively. However, in adult farm goats and sheep using s-Kit 1, 25.0 and 43.7% animals were positives, as compared to 17.8 and 25.0% by Kit 2, respectively. In farmer's buffaloes s-Kit 1 detected 58.6% buffaloes positive, whereas, none was positive in Kit 2. In human beings, s-Kit 1 and Kit 2 had 34.0- 42.1 and 30.0-40.7%, positives, respectively, Comparative evaluation of 3 kits on 72 serum samples of farm goats and sheep showed that sensitivity and specificity were, 55.5 and 86.3 and 18.5 and 86.5 and 3.7 and 91.7% in Kit 1, 2 and 3, respectively. S/P ratios showed that Kit 1 in comparison to culture (fecal, tissues, milk) was never over sensitive. Indigenous ELISA kit (Kit 1), was a useful 'Multi-Species kit' for screening of MAP infection in animals and human beings in India.