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Ready-to-use qPCR for detection of Cyclospora cayetanensis or Trypanosoma cruzi in food matrices.
Costa, Alexandre D T; Jacomasso, Thiago; Mattos, Elaine C; Farias, Aline B; Rampazzo, Rita C P; Pinto, Rebeka S; Tassi, Walleyd; Marciano, Maria Aparecida M; Pereira-Chioccola, Vera Lucia; Murphy, Helen R; da Silva, Alexandre J; Krieger, Marco A.
Afiliación
  • Costa ADT; Laboratório de Ciências e Tecnologias Aplicadas à Saúde (LaCTAS), Instituto Carlos Chagas (ICC), Fundação Oswaldo Cruz, Rua Professor Algacyr Munhoz Mader 3775, Curitiba 81350-010, Brazil.
  • Jacomasso T; Instituto de Biologia Molecular do Paraná (IBMP), Rua Professor Algacyr Munhoz Mader 3775, Curitiba 81350-010, Brazil.
  • Mattos EC; Instituto de Biologia Molecular do Paraná (IBMP), Rua Professor Algacyr Munhoz Mader 3775, Curitiba 81350-010, Brazil.
  • Farias AB; Instituto Adolfo Lutz (IAL Santo André), Av. Ramiro Colleone 240, Santo André, SP 09040-160, Brazil.
  • Rampazzo RCP; Instituto de Biologia Molecular do Paraná (IBMP), Rua Professor Algacyr Munhoz Mader 3775, Curitiba 81350-010, Brazil.
  • Pinto RS; Instituto de Biologia Molecular do Paraná (IBMP), Rua Professor Algacyr Munhoz Mader 3775, Curitiba 81350-010, Brazil.
  • Tassi W; Instituto de Biologia Molecular do Paraná (IBMP), Rua Professor Algacyr Munhoz Mader 3775, Curitiba 81350-010, Brazil.
  • Marciano MAM; Instituto de Biologia Molecular do Paraná (IBMP), Rua Professor Algacyr Munhoz Mader 3775, Curitiba 81350-010, Brazil.
  • Pereira-Chioccola VL; Instituto Adolfo Lutz (IAL Central), Av. Dr. Arnaldo 355, São Paulo, SP 01246-000, Brazil.
  • Murphy HR; Instituto Adolfo Lutz (IAL Central), Av. Dr. Arnaldo 355, São Paulo, SP 01246-000, Brazil.
  • da Silva AJ; U.S. Food & Drug Administration Center for Food Safety and Applied Nutrition (CFSAN), Office of Applied Research and Safety Assessment, 8301 Muirkirk Road, Laurel, MD 21403, USA.
  • Krieger MA; U.S. Food & Drug Administration Center for Food Safety and Applied Nutrition (CFSAN), Office of Applied Research and Safety Assessment, 8301 Muirkirk Road, Laurel, MD 21403, USA.
Food Waterborne Parasitol ; 22: e00111, 2021 Mar.
Article en En | MEDLINE | ID: mdl-33681489
Foodborne outbreaks caused by parasites have long been a public health issue. Among the available contamination detection methods, qPCR is one of the most sensitive and specific. However, it can be cumbersome and error-prone, if used by unexperienced users. Moreover, qPCR reagents usually require freezer temperatures for transportation and storage. We present a gelified reaction format that allows the reagents to be stored at 2-8 °C for up to 90 days without losing performance. The gelification process eliminates most operator mistakes during reaction setup, and renders the qPCR plates ready-to-use. The new reaction makeup was evaluated using artificially contaminated samples of distinct food matrices for sensitivity, specificity, repeatability, reproducibility, and stability. Samples consisted of cilantro leaves and raspberry fruits spiked with Cyclospora cayetanensis oocysts, as well as açai pulp and sugarcane juice tainted with Trypanosoma cruzi trypomastigotes. No significant difference between the gelified and the non-gelified qPCR was found. Our results suggest that gelifying the assay may help to achieve more reproducible qPCR data across laboratories, thus supporting surveillance actions. (170 words).
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Tipo de estudio: Diagnostic_studies Idioma: En Revista: Food Waterborne Parasitol Año: 2021 Tipo del documento: Article País de afiliación: Brasil Pais de publicación: Países Bajos
Texto completo
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XML
PubMed Links
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Tipo de estudio: Diagnostic_studies Idioma: En Revista: Food Waterborne Parasitol Año: 2021 Tipo del documento: Article País de afiliación: Brasil Pais de publicación: Países Bajos