The present invention concerns a composition of polypeptides suitable for detecting antibodies against Trypanosoma cruzi (T. cruzi) in an isolated biological sample consisting of three polypeptides 1F8, JL7 and Cruzipain. A method of producing a soluble and immunoreactive composition of polypeptides suitable for detecting antibodies against T. cruzi using said composition of polypeptides is also part of the invention. Moreover, the invention concerns a method for detecting antibodies specific for T. cruzi in an isolated sample wherein a composition of said T. cruzi polypeptides is used as well as a reagent kit comprising said composition of T. cruzi polypeptides.