Interlaboratory validation of an improved method for detection of Cyclospora cayetanensis in produce using a real-time PCR assay
| Autor: | Kristopher J. Stanya, Jason Neal-McKinney, June H. Wetherington, Fernanda S. Nascimento, Lacresha D. Chatman, Gabrielle S. Pires, Mansour Samadpour, Amy M. Kahler, Hediye Nese Cinar, Kathy E. Noe, Gopal R. Gopinath, Aleksey Molokin, Cynthia L. Johnson, Helen R. Murphy, Mónica Santín, Harish K Janagama, Candace Miller, Elizabeth Sachs, Nancy Miranda, Alexandre J. da Silva |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Validation study 030106 microbiology Coriandrum Food Contamination Improved method Biology Real-Time Polymerase Chain Reaction DNA Ribosomal Microbiology Cyclospora cayetanensis Food and drug administration 03 medical and health sciences Sample preparation 18s rdna Chromatography United States Food and Drug Administration Reproducibility of Results biology.organism_classification Protozoan parasite United States Cyclospora 030104 developmental biology Real-time polymerase chain reaction Rubus Food Science |
| Zdroj: | Food Microbiology. 69:170-178 |
| ISSN: | 0740-0020 |
| DOI: | 10.1016/j.fm.2017.08.008 |
| Popis: | A collaborative validation study was performed to evaluate the performance of a new U.S. Food and Drug Administration method developed for detection of the protozoan parasite, Cyclospora cayetanensis, on cilantro and raspberries. The method includes a sample preparation step in which oocysts are recovered from produce using an enhanced produce washing solution containing 0.1% Alconox and a commercially available method to disrupt the C. cayetanensis oocysts and extract DNA. A real-time PCR assay targeting the C. cayetanensis 18S rDNA gene with an internal amplification control to monitor PCR inhibition provides species-specific identification. Five laboratories blindly analyzed a total of 319 samples consisting of 25 g of cilantro or 50 g of raspberries which were either uninoculated or artificially contaminated with C. cayetanensis oocysts. Detection rates for cilantro inoculated with 200, 10, and 5 oocysts, were 100%, 80%, and 31%, respectively. For raspberries, the detection rates for samples inoculated with 200, 10, and 5 oocysts were 100%, 90% and 50%, respectively. All uninoculated samples, DNA blank extracts, and no-template PCR controls were negative. Reproducibility between laboratories and analysts was high and the method was shown to be an effective analytical tool for detection of C. cayetanensis in produce. |
| Databáze: | OpenAIRE |
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