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ABSTRACT. de Moraes E.P.B.X., Freitas A.C., Costa M.M., PinheiroJr J.W. & Mota R.A. [Nested PCR utilization for Toxoplasma gondii detection in the blood of experimentally infected sheep]. Utilização da PCR nested para detecção de Toxoplasma gondii no sangue de ovelhas experimentalmente infectadas. Revista Brasileira de Medicina Veterinária, 35(4):329-334, 2013. Laboratório de Doenças Infecto-Contagiosas dos Animais Domésticos, Departamento de Medicina Veterinária, Universidade Federal Rural de Pernambuco, Rua Dom Manoel de Medeiros, s/n, Dois Irmãos, Recife, PE 52171-900, Brasil. E-mail: firstname.lastname@example.org The aim of this study was to evaluate the sensitivity of the PCR and nested PCR for T. gondii detection in the blood of experimentally infected sheep with tachyzoites through the semen. Blood samples of 41 sheep were taken, which were infected with different dosages: G1 - 6.5 X 104 tachyzoites; G2- 4 X 107 tachyzoites and G3- control group. For the anti-T.gondii antibodies research the Indirect Immunofluorescence (IIF) technique was used. For the PCR and nested PCR, primers derived from B1 gene were used. Seroconversion was observed only for the infected group. In the PCR, T. gondii DNA was detected in 40% of the infected sheep blood samples, and in the nested PCR, in 93.3% of the samples. In the G3, no antibodies were detected, as well as parasitic DNA in neither of the animals. It is possible to conclude that the nested PCR is more sensitive for the T. gondii detection in the blood of animals experimentally infected through the semen, presenting superior results in comparison to those of the PCR and might be successfully used in studies like this one.