{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,12]],"date-time":"2026-09-12T18:53:35Z","timestamp":1789239215757,"version":"build-2803163510"},"reference-count":54,"publisher":"Elsevier BV","license":[{"start":{"date-parts":[[2025,6,1]],"date-time":"2025-06-01T00:00:00Z","timestamp":1748736000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/www.elsevier.com\/tdm\/userlicense\/1.0\/"},{"start":{"date-parts":[[2025,6,1]],"date-time":"2025-06-01T00:00:00Z","timestamp":1748736000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/www.elsevier.com\/legal\/tdmrep-license"},{"start":{"date-parts":[[2025,1,8]],"date-time":"2025-01-08T00:00:00Z","timestamp":1736294400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100021250","name":"US Food and Drug Administration Human Foods Program","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100021250","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["elsevier.com","sciencedirect.com"],"crossmark-restriction":true},"short-container-title":["Food Microbiology"],"published-print":{"date-parts":[[2025,6]]},"DOI":"10.1016\/j.fm.2025.104727","type":"journal-article","created":{"date-parts":[[2025,1,9]],"date-time":"2025-01-09T00:00:14Z","timestamp":1736380814000},"page":"104727","update-policy":"https:\/\/doi.org\/10.1016\/elsevier_cm_policy","source":"Crossref","is-referenced-by-count":8,"special_numbering":"C","title":["Multi-laboratory validation of a modified real-time PCR assay (Mit1C) for the detection of Cyclospora cayetanensis in fresh produce"],"prefix":"10.1016","volume":"128","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0558-5488","authenticated-orcid":false,"given":"Sonia","family":"Almeria","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John","family":"Grocholl","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeremi","family":"Mullins","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mauricio","family":"Durigan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Laura","family":"Ewing-Peeples","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ellie Lauren","family":"Rogers","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kirsten","family":"Hirneisen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shauna","family":"Madson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shizhen Steven","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"78","reference":[{"key":"10.1016\/j.fm.2025.104727_bib1","doi-asserted-by":"crossref","first-page":"281","DOI":"10.3390\/microorganisms12020281","article-title":"Editorial for the special issue Cyclospora cayetanensis and cyclosporiasis","volume":"12","author":"Almeria","year":"2024","journal-title":"Microorganisms"},{"key":"10.1016\/j.fm.2025.104727_bib2","doi-asserted-by":"crossref","DOI":"10.1016\/j.fm.2021.103802","article-title":"Detection of Cyclospora cayetanensis on bagged pre-cut salad mixes within their shelf-life and after sell by date by the U.S. Food and Drug administration validated method","volume":"98","author":"Almeria","year":"2021","journal-title":"Food Microbiol."},{"key":"10.1016\/j.fm.2025.104727_bib3","doi-asserted-by":"crossref","first-page":"497","DOI":"10.1016\/j.fm.2018.07.013","article-title":"Evaluation of the U.S. Food and Drug Administration validated method for detection of Cyclospora cayetanensis in high-risk fresh produce matrices and a method modification for a prepared dish","volume":"76","author":"Almeria","year":"2018","journal-title":"Food Microbiol."},{"issue":"9","key":"10.1016\/j.fm.2025.104727_bib4","doi-asserted-by":"crossref","first-page":"317","DOI":"10.3390\/microorganisms7090317","article-title":"Cyclospora cayetanensis and cyclosporiasis: an update","volume":"7","author":"Almeria","year":"2019","journal-title":"Microorganisms"},{"key":"10.1016\/j.fm.2025.104727_bib5","doi-asserted-by":"crossref","DOI":"10.1016\/j.fm.2020.103719","article-title":"Modifications of the U.S. food and drug administration validated method for detection of Cyclospora cayetanensis oocysts in prepared dishes: Mexican-style salsas and guacamole","volume":"96","author":"Almeria","year":"2021","journal-title":"Food Microbiol."},{"key":"10.1016\/j.fm.2025.104727_bib6","doi-asserted-by":"crossref","first-page":"2171","DOI":"10.3390\/microorganisms11092171","article-title":"Cyclospora cayetanensis: a perspective (2020-2023) with emphasis on epidemiology and detection methods","volume":"11","author":"Almeria","year":"2023","journal-title":"Microorganisms"},{"key":"10.1016\/j.fm.2025.104727_bib7","doi-asserted-by":"crossref","first-page":"1506","DOI":"10.3390\/microorganisms11061506","article-title":"Molecular detection of Cyclospora cayetanensis in two main types of farm soil using real-time PCR assays and method modification for commercial potting mix","volume":"11","author":"Arida","year":"2023","journal-title":"Microorganisms"},{"key":"10.1016\/j.fm.2025.104727_bib8","doi-asserted-by":"crossref","DOI":"10.1016\/j.fm.2019.103397","article-title":"Evaluation of the U.S. Food and Drug Administration validated molecular method for detection of Cyclospora cayetanensis oocysts on fresh and frozen berries","volume":"87","author":"Assurian","year":"2020","journal-title":"Food Microbiol."},{"key":"10.1016\/j.fm.2025.104727_bib9","doi-asserted-by":"crossref","DOI":"10.1016\/j.foodcont.2021.108656","article-title":"Global prevalence of intestinal protozoan contamination in vegetables and fruits: a systematic review and meta-analysis","volume":"133","author":"Badri","year":"2022","journal-title":"Food Control"},{"key":"10.1016\/j.fm.2025.104727_bib26","series-title":"FDA Guidelines for the Validation of Microbiological Methods for the FDA Foods Program","year":"2024"},{"key":"10.1016\/j.fm.2025.104727_bib11","unstructured":"BAM Chapter 19b: Molecular Detection of Cyclospora Cayetanensis in Fresh Produce Using Real-Time PCR | FDA) (accessed October 2024)."},{"key":"10.1016\/j.fm.2025.104727_bib10","doi-asserted-by":"crossref","DOI":"10.1016\/j.jfp.2022.100037","article-title":"Development and single laboratory evaluation of a refined and specific real-time PCR detection method, using mitochondrial primers (Mit1C), for the detection of Cyclospora cayetanensis in produce","volume":"86","author":"Balan","year":"2023","journal-title":"J. Food Protect."},{"key":"10.1016\/j.fm.2025.104727_bib12","doi-asserted-by":"crossref","DOI":"10.1016\/j.fm.2021.103792","article-title":"Contamination of fresh produce sold on the Italian market with Cyclospora cayetanensis and Echinococcus multilocularis","author":"Barlaam","year":"2021","journal-title":"Food Microbiol."},{"key":"10.1016\/j.fm.2025.104727_bib13","article-title":"Detection of Cyclospora cayetanensis, Echinococcus multilocularis, Toxocara spp. and microsporidia in fresh produce using molecular methods\u2014a review","volume":"23","author":"Bartosova","year":"2021","journal-title":"Food Waterborne Parasitol"},{"key":"10.1016\/j.fm.2025.104727_bib14","doi-asserted-by":"crossref","first-page":"611","DOI":"10.1373\/clinchem.2008.112797","article-title":"The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments","volume":"55","author":"Bustin","year":"2009","journal-title":"Clin. Chem."},{"key":"10.1016\/j.fm.2025.104727_bib15","doi-asserted-by":"crossref","first-page":"1101","DOI":"10.15585\/mmwr.mm6739a6","article-title":"Multiple cyclosporiasis outbreaks\u2014United States, 2018","volume":"67","author":"Casillas","year":"2018","journal-title":"MMWR (Morb. Mortal. Wkly. Rep.)"},{"key":"10.1016\/j.fm.2025.104727_bib16","series-title":"Domestically Acquired Cases of Cyclosporiasis \u2014 United States, May\u2013August 2022. Cyclosporiasis Outbreak Investigations \u2014 United States, 2022 (cdc.Gov)","year":"2022"},{"key":"10.1016\/j.fm.2025.104727_bib17","doi-asserted-by":"crossref","first-page":"673","DOI":"10.1093\/trstmh\/trab179","article-title":"Seasonal trends of Cyclospora cayetanensis infections in a community of falcon state, northwestern Venezuela","volume":"116","author":"Chacin-Bonilla","year":"2022","journal-title":"Trans. R. Soc. Trop. Med. Hyg."},{"key":"10.1016\/j.fm.2025.104727_bib18","doi-asserted-by":"crossref","first-page":"1348","DOI":"10.4315\/0362-028X.JFP-13-381","article-title":"Efficacy of wash solutions in recovering Cyclospora cayetanensis, Cryptosporidium parvum, and Toxoplasma gondii from basil","volume":"77","author":"Chandra","year":"2014","journal-title":"J. Food Protect."},{"key":"10.1016\/j.fm.2025.104727_bib19","doi-asserted-by":"crossref","DOI":"10.1016\/j.actatropica.2024.107175","article-title":"The global prevalence of Cyclospora cayetanensis infection: a systematic review, meta-analysis, and meta-regression","volume":"253","author":"Chen","year":"2024","journal-title":"Acta Trop."},{"key":"10.1016\/j.fm.2025.104727_bib20","doi-asserted-by":"crossref","first-page":"1216","DOI":"10.1086\/514780","article-title":"Cyclosporiasis: clinical and histopathologic correlates","volume":"28","author":"Connor","year":"1999","journal-title":"Clin. Infect. Dis."},{"key":"10.1016\/j.fm.2025.104727_bib21","doi-asserted-by":"crossref","DOI":"10.1016\/j.fm.2023.104299","article-title":"Multi-laboratory validation study of a real-time PCR method for detection of Salmonella in baby spinach","volume":"114","author":"Deng","year":"2023","journal-title":"Food Microbiol."},{"key":"10.1016\/j.fm.2025.104727_bib22","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1016\/j.cofs.2016.04.009","article-title":"Parasitic illnesses associated with the consumption of fresh produce\u2014an emerging issue in developed countries","volume":"8","author":"Dixon","year":"2016","journal-title":"Curr. Opin. Food Sci."},{"key":"10.1016\/j.fm.2025.104727_bib23","doi-asserted-by":"crossref","DOI":"10.1016\/j.jfp.2024.100291","article-title":"Detection of Cyclospora cayetanensis in food and water samples: optimized protocols for specific and sensitive molecular methods from a regulatory agency perspective","volume":"87","author":"Durigan","year":"2024","journal-title":"J. Food Protect."},{"key":"10.1016\/j.fm.2025.104727_bib24","first-page":"2021","article-title":"FDA Releases Cyclospora prevention, response, and research action plan","year":"2021","journal-title":"FDA Food and Drug Administration"},{"key":"10.1016\/j.fm.2025.104727_bib25","unstructured":"FDA Foods Program Compendium of Analytical Laboratory Methods: Microbiological Methods (Foods Program Compendium of Analytical Laboratory Methods | FDA) (accessed December 2024)."},{"key":"10.1016\/j.fm.2025.104727_bib27","doi-asserted-by":"crossref","first-page":"1040","DOI":"10.1086\/314051","article-title":"Cyclospora cayetanensis: a review, focusing on the outbreaks of cyclosporiasis in the 1990s","volume":"31","author":"Herwaldt","year":"2000","journal-title":"Clin. Infect. Dis."},{"key":"10.1016\/j.fm.2025.104727_bib28","article-title":"Microbiology of the food chain \u2014 method validation \u2014 Part 2: protocol for the validation of alternative (proprietary) methods against a reference method","year":"2016","journal-title":"Link: ISO 16140-2:2016(en), Microbiology of the food chain \u2014 Method validation \u2014 Part 2: Protocol for the validation of alternative (proprietary) methods against a reference method"},{"key":"10.1016\/j.fm.2025.104727_bib29","unstructured":"ISO 18744:2016\/CD- Amd.1(E). Microbiology of the food chain \u2014 detection of Cryptosporidium and Giardia on fresh leafy green vegetables and berry fruits. Amendment 1: Method Validation Studies and Performance Characteristics."},{"key":"10.1016\/j.fm.2025.104727_bib30","series-title":"Public Health Risks Associated with Food-Borne Parasites. Efsa J","author":"Koutsoumanis","year":"2018"},{"key":"10.1016\/j.fm.2025.104727_bib31","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.fawpar.2015.12.002","article-title":"Optimization and validation of methods for isolation and real-time PCR identification of protozoan oocysts on leafy green vegetables and berry fruits","volume":"2","author":"Lalonde","year":"2016","journal-title":"Food Waterborne Parasitol."},{"key":"10.1016\/j.fm.2025.104727_bib32","doi-asserted-by":"crossref","first-page":"559","DOI":"10.3390\/microorganisms10030559","article-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","volume":"10","author":"Lalonde","year":"2022","journal-title":"Microorganisms"},{"key":"10.1016\/j.fm.2025.104727_bib33","doi-asserted-by":"crossref","first-page":"43","DOI":"10.3389\/fcimb.2020.00043","article-title":"Advances in cyclosporiasis diagnosis and therapeutic Intervention","volume":"10","author":"Li","year":"2020","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"10.1016\/j.fm.2025.104727_bib34","doi-asserted-by":"crossref","first-page":"1086","DOI":"10.5740\/jaoacint.12-377","article-title":"Determination of the relative level of detection of a Qualitative microbiological measurement method with respect to a reference measurement method","volume":"96","author":"M\u0103rg\u0103ritescu","year":"2013","journal-title":"J. AOAC Int."},{"key":"10.1016\/j.fm.2025.104727_bib35","doi-asserted-by":"crossref","DOI":"10.2807\/1560-7917.ES.2017.22.32.30592","article-title":"Cyclosporiasis in travellers returning to the United Kingdom from Mexico in summer 2017: lessons from the recent past to inform the future","volume":"22","author":"Marques","year":"2017","journal-title":"Euro Surveill."},{"key":"10.1016\/j.fm.2025.104727_bib36","doi-asserted-by":"crossref","first-page":"1863","DOI":"10.3390\/microorganisms9091863","article-title":"Cyclosporiasis\u2014updates on clinical presentation, pathology, clinical diagnosis, and treatment","volume":"9","author":"Mathison","year":"2021","journal-title":"Microorganisms"},{"key":"10.1016\/j.fm.2025.104727_bib37","doi-asserted-by":"crossref","first-page":"225","DOI":"10.14745\/ccdr.v45i09a01","article-title":"Use of a case-control study and control bank to investigate an outbreak of locally acquired cyclosporiasis in Canada, 2016","volume":"45","author":"Morton","year":"2019","journal-title":"Can. Comm. Dis. Rep."},{"key":"10.1016\/j.fm.2025.104727_bib38","doi-asserted-by":"crossref","first-page":"170","DOI":"10.1016\/j.fm.2017.08.008","article-title":"Interlaboratory validation of an improved method for detection of Cyclospora cayetanensis in produce using a real-time PCR assay","volume":"69","author":"Murphy","year":"2018","journal-title":"Food Microbiol."},{"key":"10.1016\/j.fm.2025.104727_bib39","doi-asserted-by":"crossref","first-page":"537","DOI":"10.4269\/ajtmh.13-0535","article-title":"Cyclospora cayetanensis in a pediatric hospital in Morelia, M\u00e9xico","volume":"91","author":"Orozco-Mosqueda","year":"2014","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"10.1016\/j.fm.2025.104727_bib40","doi-asserted-by":"crossref","first-page":"218","DOI":"10.1128\/CMR.00026-09","article-title":"Update on Cyclospora cayetanensis, a food-borne and waterborne parasite","volume":"23","author":"Ortega","year":"2010","journal-title":"Clin. Microbiol. Rev."},{"key":"10.1016\/j.fm.2025.104727_bib41","doi-asserted-by":"crossref","first-page":"1679","DOI":"10.4315\/JFP-20-121","article-title":"Multiplex quantitative PCR analysis of strawberries from Bogot\u00e1, Colombia, for contamination with three parasites","volume":"83","author":"Ortiz-Pineda","year":"2020","journal-title":"J. Food Protect."},{"key":"10.1016\/j.fm.2025.104727_bib42","series-title":"R: A Language and Environment for Statistical Computing","year":"2023"},{"key":"10.1016\/j.fm.2025.104727_bib43","doi-asserted-by":"crossref","first-page":"261","DOI":"10.3390\/foods9030261","article-title":"A molecular tool for rapid detection and traceability of Cyclospora cayetanensis in fresh berries and berry farm soils","volume":"9","author":"Resendiz-Nava","year":"2020","journal-title":"Foods"},{"key":"10.1016\/j.fm.2025.104727_bib44","doi-asserted-by":"crossref","DOI":"10.1128\/spectrum.01388-23","article-title":"Comparison of two novel one-tube nested real-time qPCR assays to detect human-infecting Cyclospora spp","volume":"11","author":"Richins","year":"2023","journal-title":"Microbiol. Spectr."},{"key":"10.1016\/j.fm.2025.104727_bib45","first-page":"39e44","article-title":"Real-time PCR in food science: PCR diagnostics","volume":"15","author":"Rodriguez-Lazaro","year":"2013","journal-title":"Curr. Issues Mol. Biol."},{"key":"10.1016\/j.fm.2025.104727_bib46","doi-asserted-by":"crossref","DOI":"10.1016\/j.fm.2019.103252","article-title":"Simultaneous detection of four protozoan parasites on leafy greens using a novel multiplex PCR assay","volume":"84","author":"Shapiro","year":"2019","journal-title":"Food Microbiol."},{"key":"10.1016\/j.fm.2025.104727_bib47","doi-asserted-by":"crossref","first-page":"167","DOI":"10.3390\/microorganisms9010167","article-title":"Molecular methods for the detection of Toxoplasma gondii oocysts in fresh produce: an extensive review","volume":"9","author":"Slana","year":"2021","journal-title":"Microorganisms"},{"key":"10.1016\/j.fm.2025.104727_bib48","doi-asserted-by":"crossref","first-page":"277","DOI":"10.1016\/S0167-7012(03)00036-8","article-title":"Sensitivity of PCR detection of Cyclospora cayetanensis in raspberries, basil, and mesclun lettuce","volume":"54","author":"Steele","year":"2003","journal-title":"J. Microbiol. Methods"},{"key":"10.1016\/j.fm.2025.104727_bib49","doi-asserted-by":"crossref","first-page":"1939","DOI":"10.3389\/fmicb.2019.01939","article-title":"A new protocol for molecular detection of Cyclospora cayetanensis as contaminants of berry fruits","volume":"10","author":"Temesgen","year":"2019","journal-title":"Front. Microbiol."},{"key":"10.1016\/j.fm.2025.104727_bib50","doi-asserted-by":"crossref","DOI":"10.1016\/j.fm.2022.103980","article-title":"Surveillance of berries sold on the Norwegian market for parasite contamination using molecular methods","volume":"104","author":"Temesgen","year":"2022","journal-title":"Food Microbiol."},{"key":"10.1016\/j.fm.2025.104727_bib51","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1078\/1438-4221-00252","article-title":"Detection of Cyclospora cayetanensis in travelers returning from the tropics and subtropics using microscopy and real-time PCR","volume":"293","author":"Verweij","year":"2003","journal-title":"Int. J. Med. Microbiol."},{"key":"10.1016\/j.fm.2025.104727_bib52","article-title":"On the estimation of POD and LOD of qualitative microbiological assays from a multi-laboratory validation study","author":"Wang","year":"2021","journal-title":"J. AOAC Int."},{"key":"10.1016\/j.fm.2025.104727_bib53","doi-asserted-by":"crossref","first-page":"1","DOI":"10.18637\/jss.v016.i09","article-title":"Object-oriented computation of sandwich estimators","volume":"16","author":"Zeileis","year":"2006","journal-title":"J. Stat. Software"},{"key":"10.1016\/j.fm.2025.104727_bib54","first-page":"1","article-title":"Various versatile variances: an object-oriented implementation of clustered covariances in R. J","volume":"95","author":"Zeileis","year":"2020","journal-title":"Statistical Software"}],"container-title":["Food Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.elsevier.com\/content\/article\/PII:S0740002025000073?httpAccept=text\/xml","content-type":"text\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/api.elsevier.com\/content\/article\/PII:S0740002025000073?httpAccept=text\/plain","content-type":"text\/plain","content-version":"vor","intended-application":"text-mining"}],"deposited":{"date-parts":[[2025,2,23]],"date-time":"2025-02-23T21:29:29Z","timestamp":1740346169000},"score":1,"resource":{"primary":{"URL":"https:\/\/linkinghub.elsevier.com\/retrieve\/pii\/S0740002025000073"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,6]]},"references-count":54,"alternative-id":["S0740002025000073"],"URL":"https:\/\/doi.org\/10.1016\/j.fm.2025.104727","relation":{},"ISSN":["0740-0020"],"issn-type":[{"value":"0740-0020","type":"print"}],"subject":[],"published":{"date-parts":[[2025,6]]},"assertion":[{"value":"Elsevier","name":"publisher","label":"This article is maintained by"},{"value":"Multi-laboratory validation of a modified real-time PCR assay (Mit1C) for the detection of Cyclospora cayetanensis in fresh produce","name":"articletitle","label":"Article Title"},{"value":"Food Microbiology","name":"journaltitle","label":"Journal Title"},{"value":"https:\/\/doi.org\/10.1016\/j.fm.2025.104727","name":"articlelink","label":"CrossRef DOI link to publisher maintained version"},{"value":"article","name":"content_type","label":"Content Type"},{"value":"Published by Elsevier Ltd.","name":"copyright","label":"Copyright"}],"article-number":"104727"}}