Bovine trypanosomiasis requires, even in situations of low parasitemia, a rapid, sensitive and specific diagnosis in order to take the correct therapeutic measures and avoid the dissemination in herds and significant economic losses. Therefore, the study was designed to evaluate the comparative performance of molecular assays PCR, real-time PCR and LAMP in the detection of Trypanosoma vivax in blood samples from cattle and buffalo naturally infected in the Brazilian Amazon. PCR indicated a 4% (06/150) prevalence of the protozoan parasite in the tested herds while real-time PCR and LAMP estimated 38% (57/150) and 83.33% (125/150) of animals with trypanosomiasis, respectively. The kappa analysis denoted reasonable agreement between the results of real-time PCR and LAMP and poor agreement between conventional PCR and the other techniques, as interpreted by Landis and koch, 1977. Despite this, the higher percentage of parasite detection obtained by the LAMP assay coincides with its reputation as a highly sensitive method, suggesting that it is a viable alternative to traditional molecular techniques with potential use in epidemiological screening and clinical diagnostic efficiency. However, further studies are required to adjust LAMP protocols to facilitate the implementation and reading of the results, ensuring their applicability in the laboratorial routine.