Trypanosoma cruzi: A new system for primary culture and isolation of the parasite.

Various methods, including blood culture, are employed to isolate Trypanosoma cruzi. However, there is currently no standardized protocol for parasite culture, and the effectiveness of available techniques varies. In this study, we developed a standardized closed blood culture system (CBCS) designed to support the survival of trypomastigotes and their differentiation into epimastigotes, from sample collection through to primary laboratory isolation. Blood samples were artificially infected with varying concentrations of T. cruzi trypomastigotes to assess the performance of the CBCS. The CBCS enabled successful isolation and exponential growth of the parasite, demonstrating performance comparable to that of conventional culture methods. All assays included a reference control culture, which served as a benchmark for comparison. No contamination events were observed, and it was possible to isolate and expand the parasite population from an initial sample containing as few as ten trypomastigotes. The standardized CBCS protocol demonstrated good precision, as confirmed by repeatability and reproducibility tests, which showed acceptable variability among replicates.