title: Extraction of Trypanosoma cruzi DNA from food: a contribution to the elucidation of acute Chagas disease outbreaks.
abstract: BACKGROUND
  Before 2004, the occurrence of acute Chagas disease (ACD) by oral transmission associated with food was scarcely known or investigated. Originally sporadic and circumstantial, ACD occurrences have now become frequent in the Amazon region, with recently related outbreaks spreading to several Brazilian states. These cases are associated with the consumption of açai juice by waste reservoir animals or insect vectors infected with Trypanosoma cruzi in endemic areas. Although guidelines for processing the fruit to minimize contamination through microorganisms and parasites exist, açai-based products must be assessed for quality, for which the demand for appropriate methodologies must be met.
  
  
  METHODS
  Dilutions ranging from 5 to 1,000 T. cruzi CL Brener cells were mixed with 2mL of acai juice. Four Extraction of T. cruzi DNA methods were used on the fruit, and the cetyltrimethyl ammonium bromide (CTAB) method was selected according to JRC, 2005.
  
  
  RESULTS
  DNA extraction by the CTAB method yielded satisfactory results with regard to purity and concentration for use in PCR. Overall, the methods employed proved that not only extraction efficiency but also high sensitivity in amplification was important.
  
  
  CONCLUSIONS
  The method for T. cruzi detection in food is a powerful tool in the epidemiological investigation of outbreaks as it turns epidemiological evidence into supporting data that serve to confirm T. cruzi infection in the foods. It also facilitates food quality control and assessment of good manufacturing practices involving acai-based products.
authors.affiliation: Programa de Pós-Graduação em Vigilância Sanitária, Instituto Nacional de Controle de Qualidade em Saúde, Fundação Oswaldo Cruz, Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Renata Trotta Barroso
authors.initials: RT
authors.lastname: Ferreira
authors.affiliation: Instituto Nacional de Controle de Qualidade em Saúde, Fundação Oswaldo Cruz, Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Aline Martins
authors.initials: AM
authors.lastname: Melandre
authors.affiliation: Instituto Nacional de Controle de Qualidade em Saúde, Fundação Oswaldo Cruz, Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Maria Luiza
authors.initials: ML
authors.lastname: Cabral
authors.affiliation: Instituto Nacional de Controle de Qualidade em Saúde, Fundação Oswaldo Cruz, Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Maria Regina
authors.initials: MR
authors.lastname: Branquinho
authors.affiliation: Instituto Nacional de Controle de Qualidade em Saúde, Fundação Oswaldo Cruz, Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Paola
authors.initials: P
authors.lastname: Cardarelli-Leite
mesh_terms: D000818:Animals; D014355:Chagas Disease / Q000453:epidemiology / Q000635:transmission*; D016054:DNA, Protozoan / Q000302:isolation & purification*; D004196:Disease Outbreaks; D065748:Euterpe / Q000469:parasitology*; D005506:Food Contamination; D016266:Food Parasitology; D006801:Humans; D016133:Polymerase Chain Reaction; D014349:Trypanosoma cruzi / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D016054:DNA, Protozoan