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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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Detalles Bibliográficos
Autores principales: Lalonde, Laura, Oakley, Jenna, Fries, Patrick
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
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
Descripción
Sumario: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.