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Verification and Use of the US-FDA BAM 19b Method for Detection of Cyclospora cayetanensis in a Survey of Fresh Produce by CFIA Laboratory
To facilitate the harmonized surveillance and investigation of cyclosporiasis outbreaks in the US and Canada, we adapted and verified the US-FDA’s BAM 19b method and employed it in a national produce survey. Performance was verified by spiking 200, 10, 5 or 0 C. cayetanensis oocysts onto berries (50...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8954584/ https://www.ncbi.nlm.nih.gov/pubmed/35336134 http://dx.doi.org/10.3390/microorganisms10030559 |
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author | Lalonde, Laura Oakley, Jenna Fries, Patrick |
author_facet | Lalonde, Laura Oakley, Jenna Fries, Patrick |
author_sort | Lalonde, Laura |
collection | PubMed |
description | To facilitate the harmonized surveillance and investigation of cyclosporiasis outbreaks in the US and Canada, we adapted and verified the US-FDA’s BAM 19b method and employed it in a national produce survey. Performance was verified by spiking 200, 10, 5 or 0 C. cayetanensis oocysts onto berries (50 ± 5 g, n = 85) and 200, 10 or 0 oocysts onto green onions (25 ± 3 g, n = 24) and leafy greens (25 ± 1 g, n = 120) and testing these samples by the BAM method on Bio-Rad CFX96. Method robustness was assessed by aging (0 or 7 days) and freezing the produce and washes prior to testing, then implementing the method for the surveillance testing of 1759 imported leafy green, herb and berry samples. Diagnostic sensitivity was 100/44% and 93/30% for berries and leafy greens spiked with 200/10 oocysts, respectively. The diagnostic and analytical specificity were 100% for all matrices and related parasites tested. The proportion positive was unaffected (p = 0.22) by age or condition of produce (7d, fresh, frozen) or wash concentrate (3d, fresh, frozen); however, the Cq values were higher (p = 0.009) for raspberries aged 7d (37.46 ± 0.29) compared to fresh (35.36 ± 0.29). C. cayetanensis was detected in berries (two), herbs (two) and leafy greens (one), representing 0.28% of the tested survey samples. These results independently verified the reported performance characteristics and robustness of the BAM method for the detection of C. cayetanensis in a variety of matrices, including under adverse sample conditions, using a unique detection platform and demonstrating its routine diagnostic use in our Canadian Food Inspection Agency (CFIA) laboratory. |
format | Online Article Text |
id | pubmed-8954584 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-89545842022-03-26 Verification and Use of the US-FDA BAM 19b Method for Detection of Cyclospora cayetanensis in a Survey of Fresh Produce by CFIA Laboratory Lalonde, Laura Oakley, Jenna Fries, Patrick Microorganisms Article To facilitate the harmonized surveillance and investigation of cyclosporiasis outbreaks in the US and Canada, we adapted and verified the US-FDA’s BAM 19b method and employed it in a national produce survey. Performance was verified by spiking 200, 10, 5 or 0 C. cayetanensis oocysts onto berries (50 ± 5 g, n = 85) and 200, 10 or 0 oocysts onto green onions (25 ± 3 g, n = 24) and leafy greens (25 ± 1 g, n = 120) and testing these samples by the BAM method on Bio-Rad CFX96. Method robustness was assessed by aging (0 or 7 days) and freezing the produce and washes prior to testing, then implementing the method for the surveillance testing of 1759 imported leafy green, herb and berry samples. Diagnostic sensitivity was 100/44% and 93/30% for berries and leafy greens spiked with 200/10 oocysts, respectively. The diagnostic and analytical specificity were 100% for all matrices and related parasites tested. The proportion positive was unaffected (p = 0.22) by age or condition of produce (7d, fresh, frozen) or wash concentrate (3d, fresh, frozen); however, the Cq values were higher (p = 0.009) for raspberries aged 7d (37.46 ± 0.29) compared to fresh (35.36 ± 0.29). C. cayetanensis was detected in berries (two), herbs (two) and leafy greens (one), representing 0.28% of the tested survey samples. These results independently verified the reported performance characteristics and robustness of the BAM method for the detection of C. cayetanensis in a variety of matrices, including under adverse sample conditions, using a unique detection platform and demonstrating its routine diagnostic use in our Canadian Food Inspection Agency (CFIA) laboratory. MDPI 2022-03-04 /pmc/articles/PMC8954584/ /pubmed/35336134 http://dx.doi.org/10.3390/microorganisms10030559 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Lalonde, Laura Oakley, Jenna Fries, Patrick Verification and Use of the US-FDA BAM 19b Method for Detection of Cyclospora cayetanensis in a Survey of Fresh Produce by CFIA Laboratory |
title | Verification and Use of the US-FDA BAM 19b Method for Detection of Cyclospora cayetanensis in a Survey of Fresh Produce by CFIA Laboratory |
title_full | Verification and Use of the US-FDA BAM 19b Method for Detection of Cyclospora cayetanensis in a Survey of Fresh Produce by CFIA Laboratory |
title_fullStr | Verification and Use of the US-FDA BAM 19b Method for Detection of Cyclospora cayetanensis in a Survey of Fresh Produce by CFIA Laboratory |
title_full_unstemmed | Verification and Use of the US-FDA BAM 19b Method for Detection of Cyclospora cayetanensis in a Survey of Fresh Produce by CFIA Laboratory |
title_short | Verification and Use of the US-FDA BAM 19b Method for Detection of Cyclospora cayetanensis in a Survey of Fresh Produce by CFIA Laboratory |
title_sort | verification and use of the us-fda bam 19b method for detection of cyclospora cayetanensis in a survey of fresh produce by cfia laboratory |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8954584/ https://www.ncbi.nlm.nih.gov/pubmed/35336134 http://dx.doi.org/10.3390/microorganisms10030559 |
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