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READY-TO-USE QPCR FOR DETECTION OF CYCLOSPORA CAYETANENSIS OR TRYPANOSOMA CRUZI IN FOOD MATRICES
Costa, Alexandre Dias Tavares et al. | Date Issued: 2021
Author
Costa, Alexandre Dias Tavares
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.
Silva, Alexandre J. da
Krieger, Marco Aurélio
Affilliation
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Ciências e Tecnologias Aplicadas à Saúde. Curitiba, PR, Brasil. / Fundação Oswaldo Cruz. Instituto de Biologia Molecular do Paraná. Curitiba, PR, Brasil.
Fundação Oswaldo Cruz. Instituto de Biologia Molecular do Paraná. Curitiba, PR, Brasil.
Instituto Adolfo Lutz. Santo André, SP, Brasil.
Fundação Oswaldo Cruz. Instituto de Biologia Molecular do Paraná. Curitiba, PR, Brasil.
Fundação Oswaldo Cruz. Instituto de Biologia Molecular do Paraná. Curitiba, PR, Brasil.
Fundação Oswaldo Cruz. Instituto de Biologia Molecular do Paraná. Curitiba, PR, Brasil.
Fundação Oswaldo Cruz. Instituto de Biologia Molecular do Paraná. Curitiba, PR, Brasil.
Instituto Adolfo Lutz. São Paulo, SP, Brasil.
Instituto Adolfo Lutz. São Paulo, SP, Brasil.
U.S. Food & Drug Administration Center for Food Safety and Applied Nutrition (CFSAN). Office of Applied Research and Safety Assessment. Laurel, MD, USA.
U.S. Food & Drug Administration Center for Food Safety and Applied Nutrition (CFSAN). Office of Applied Research and Safety Assessment. Laurel, MD, USA.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Ciências e Tecnologias Aplicadas à Saúde. Curitiba, PR, Brasil. / Fundação Oswaldo Cruz. Instituto de Biologia Molecular do Paraná. Curitiba, PR, Brasil.
Abstract
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.
Keywords in Portuguese
Cyclospora cayetanensis
Trypanosoma cruzi
qPCR
 
Keywords
Foodborne Diseases
Molecular Diagnostics
Protozoan Infections
Quantitative Real-Time Polymerase Chain Reaction
 
Keywords in Spanish
Enfermedades Transmitidas por los Alimentos
Patología Molecular
Infecciones por Protozoos
Reacción en Cadena en Tiempo Real de la Polimerasa
 
DeCS
Cyclospora
Doenças Transmitidas por Alimentos
Patologia Molecular
Infecções por Protozoários
Reação em Cadeia da Polimerase em Tempo Real
 
Publisher
Elsevier
Citation
COSTA, Alexandre D. T. et al. Ready-to-use qPCR for detection of Cyclospora cayetanensis or Trypanosoma cruzi in food matrices. Food and Waterborne Parasitology, n. 22, p. 1–12, 2021.
DOI
10.1016/j.fawpar.2021.e00111
ISSN
2405-6766