Comparison of DNA extraction methods and real-time PCR assays for the molecular diagnostics of chronic Chagas disease

Fil: Ramírez, Juan C. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Parasitología DR: Mario Fatala Chaben; Argentina.Fil: Silgado, Aroa. Universitat Autònoma de Barcelona, PROSICS. Hospital Universitari Vall d'Hebron. Departament de Microbiologia; Barcelona, España.Fil: Moure, Zaira. Universitat Autònoma de Barcelona, PROSICS. Hospital Universitari Vall d'Hebron. Departament de Microbiologia; España.Fil: de Oliveira, Maykon T. Universidade de São Paulo. Faculdade de Medicina de Ribeirão Preto. Unidade de Cardiologia. Departamento de Clínica Médica; Brasil.Fil: Serre-Delcor, Núria. Universitata Autònoma de Barcelona. Tropical Medicine Unit Vall d´Hebron-Drassanes; Barcelona, España.Fil: Salvador, Fernando. Universitat Autònoma de Barcelona. Department of Infectious Diseases, Vall d'Hebron University Hospital; Barcelona, España.Fil: Oliveira, Inés. Universitata Autònoma de Barcelona. Tropical Medicine Unit Vall d´Hebron-Drassanes; Barcelona, España.Fil: Molina, Israel. Universitat Autònoma de Barcelona. Department of Infectious Diseases, Vall d'Hebron University Hospital; Barcelona, España.Fil: Pumarola, Tomas. Universitat Autònoma de Barcelona, PROSICS. Hospital Universitari Vall d'Hebron. Departament de Microbiologia; España.Fil: Sulleiro, Elena. Universitat Autònoma de Barcelona, PROSICS. Hospital Universitari Vall d'Hebron. Departament de Microbiologia; España.Aim: This work aimed to compare the sensitivity of four protocols for the detection of Trypanosoma cruzi DNA in 98 blood samples from chronic Chagas disease patients. Materials & methods: Two DNA extraction (automated and manual) methods and two T. cruzi satellite DNA qPCRs (with a recent design and the usually used set of primers) were analyzed. Results: Both DNA extraction methods and qPCR assays tested in this work gave comparable qualitative results, although the lowest Ct values were obtained when samples were analyzed using the new set of primers for T. cruzi satellite DNA. Conclusion: Our results encourage the implementation of automated DNA extraction systems and the new T. cruzi qPCR for the molecular diagnostics and treatment response monitoring of chronic Chagas disease patients