Ready-to-use qPCR for detection of Cyclospora cayetanensis or Trypanosoma cruzi in food matrices
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 fr...
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doaj-9bab50b3ea42495ca062061b1a746b832021-03-03T04:22:25ZengElsevierFood and Waterborne Parasitology2405-67662021-03-0122e00111Ready-to-use qPCR for detection of Cyclospora cayetanensis or Trypanosoma cruzi in food matricesAlexandre D.T. Costa0Thiago Jacomasso1Elaine C. Mattos2Aline B. Farias3Rita C.P. Rampazzo4Rebeka S. Pinto5Walleyd Tassi6Maria Aparecida M. Marciano7Vera Lucia Pereira-Chioccola8Helen R. Murphy9Alexandre J. da Silva10Marco A. Krieger11Laboratório 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; Instituto de Biologia Molecular do Paraná (IBMP), Rua Professor Algacyr Munhoz Mader 3775, Curitiba 81350-010, Brazil; Corresponding author at: Rua Professor Algacyr Munhoz Mader 3775, Curitiba 81350-010, Brazil.Instituto de Biologia Molecular do Paraná (IBMP), Rua Professor Algacyr Munhoz Mader 3775, Curitiba 81350-010, BrazilInstituto Adolfo Lutz (IAL Santo André), Av. Ramiro Colleone 240, Santo André, SP 09040-160, BrazilInstituto de Biologia Molecular do Paraná (IBMP), Rua Professor Algacyr Munhoz Mader 3775, Curitiba 81350-010, BrazilInstituto de Biologia Molecular do Paraná (IBMP), Rua Professor Algacyr Munhoz Mader 3775, Curitiba 81350-010, BrazilInstituto de Biologia Molecular do Paraná (IBMP), Rua Professor Algacyr Munhoz Mader 3775, Curitiba 81350-010, BrazilInstituto de Biologia Molecular do Paraná (IBMP), Rua Professor Algacyr Munhoz Mader 3775, Curitiba 81350-010, BrazilInstituto Adolfo Lutz (IAL Central), Av. Dr. Arnaldo 355, São Paulo, SP 01246-000, BrazilInstituto Adolfo Lutz (IAL Central), Av. Dr. Arnaldo 355, São Paulo, SP 01246-000, BrazilU.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, USAU.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, USALaboratório 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; Instituto de Biologia Molecular do Paraná (IBMP), Rua Professor Algacyr Munhoz Mader 3775, Curitiba 81350-010, BrazilFoodborne 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)http://www.sciencedirect.com/science/article/pii/S2405676621000020Cyclospora cayetanensisTrypanosoma cruziFoodborne diseasesMolecular diagnosticProtozoanqPCR |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Alexandre D.T. Costa Thiago Jacomasso Elaine C. Mattos Aline B. Farias Rita C.P. Rampazzo Rebeka S. Pinto Walleyd Tassi Maria Aparecida M. Marciano Vera Lucia Pereira-Chioccola Helen R. Murphy Alexandre J. da Silva Marco A. Krieger |
spellingShingle |
Alexandre D.T. Costa Thiago Jacomasso Elaine C. Mattos Aline B. Farias Rita C.P. Rampazzo Rebeka S. Pinto Walleyd Tassi Maria Aparecida M. Marciano Vera Lucia Pereira-Chioccola Helen R. Murphy Alexandre J. da Silva Marco A. Krieger Ready-to-use qPCR for detection of Cyclospora cayetanensis or Trypanosoma cruzi in food matrices Food and Waterborne Parasitology Cyclospora cayetanensis Trypanosoma cruzi Foodborne diseases Molecular diagnostic Protozoan qPCR |
author_facet |
Alexandre D.T. Costa Thiago Jacomasso Elaine C. Mattos Aline B. Farias Rita C.P. Rampazzo Rebeka S. Pinto Walleyd Tassi Maria Aparecida M. Marciano Vera Lucia Pereira-Chioccola Helen R. Murphy Alexandre J. da Silva Marco A. Krieger |
author_sort |
Alexandre D.T. Costa |
title |
Ready-to-use qPCR for detection of Cyclospora cayetanensis or Trypanosoma cruzi in food matrices |
title_short |
Ready-to-use qPCR for detection of Cyclospora cayetanensis or Trypanosoma cruzi in food matrices |
title_full |
Ready-to-use qPCR for detection of Cyclospora cayetanensis or Trypanosoma cruzi in food matrices |
title_fullStr |
Ready-to-use qPCR for detection of Cyclospora cayetanensis or Trypanosoma cruzi in food matrices |
title_full_unstemmed |
Ready-to-use qPCR for detection of Cyclospora cayetanensis or Trypanosoma cruzi in food matrices |
title_sort |
ready-to-use qpcr for detection of cyclospora cayetanensis or trypanosoma cruzi in food matrices |
publisher |
Elsevier |
series |
Food and Waterborne Parasitology |
issn |
2405-6766 |
publishDate |
2021-03-01 |
description |
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) |
topic |
Cyclospora cayetanensis Trypanosoma cruzi Foodborne diseases Molecular diagnostic Protozoan qPCR |
url |
http://www.sciencedirect.com/science/article/pii/S2405676621000020 |
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