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Toxoplasma gondii is a zoonotic protozoan parasite of both medical and veterinary importance worldwide. The parasite can cause severe complications in immunocompromised individuals such as AIDS patients and transplant recipients, where up to 25% of patients will develop toxoplasmic encephalitis. Toxoplasmosis is a zoonosis that causes a public health concern in both developed and developing countries such as Thailand. Livestock development particularly in dairy cows of Thailand have been hampered by low production of milk and slow growth rate because of many pathogens including T. gondii. The objective of this study was to evaluate the serodiagnostic tool to be used for detection of T. gondii infection in dairy cows of Thailand. During 2006–2007, the sera of 700 cows from 55 small farm holders of the highest number of dairy cow population in the northern provinces were collected and analysed. Antibodies to T. gondii were assayed by latex agglutination test (LAT), enzyme-linked immunosorbent assay (ELISA) and indirect fluorescent antibody test (IFAT). The overall prevalence of T. gondii infection in dairy cows was 9.4% (66/700) by LAT and 17% (119/700) by ELISA. Sixty-three seropositive samples by LAT (95.5%) and 107 by ELISA (89.9%) were confirmed by IFAT. These results demonstrated that LAT had the highest specificity for detection of T. gondii infection. [ABSTRACT FROM AUTHOR]