{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,5]],"date-time":"2026-05-05T17:33:20Z","timestamp":1778002400446,"version":"3.51.4"},"reference-count":35,"publisher":"Public Library of Science (PLoS)","issue":"4","license":[{"start":{"date-parts":[[2025,4,25]],"date-time":"2025-04-25T00:00:00Z","timestamp":1745539200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003655","name":"National Institute of Food and Drug Safety Evaluation","doi-asserted-by":"publisher","award":["23191MFDS013"],"award-info":[{"award-number":["23191MFDS013"]}],"id":[{"id":"10.13039\/501100003655","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>Anisakidosis is a foodborne parasitic infection caused by the consumption of raw or uncooked seafood that contains third stage larvae from the Anisakidae family. This infection has been observed across the globe, with a particularly high prevalence in South Korea and Japan. Consequently, there is a necessity to compare and analyze the optimal detection methods with a view to preventing Anisakis outbreaks. In this study, a species-specific Taqman-based qPCR method was developed for the detection of the internal transcribed spacer region and mtDNA genes of <jats:italic>Anisakis simplex<\/jats:italic> and <jats:italic>Pseudoterranova decipiens<\/jats:italic>. Parasite-specific primer\/probe sets were selected based on the data from domestic and foreign detection methods. In addition, we have designed our own primer\/probe sets based on the target region of each parasite. A comprehensive literature review and a self-creation process were undertaken to select thirteen detection method sets for <jats:italic>A. simplex<\/jats:italic> and <jats:italic>P. decipiens<\/jats:italic>. The sensitivity of these sets was then evaluated by comparing the C<jats:sub>q<\/jats:sub> values from extracted DNA. The concentrations of six primer\/probe sets detected through the screening process were then compared to optimize the test method. The resultant optimized method demonstrated a limit of detection of 0.0019\u2009ng\/\u00b5L for <jats:italic>A. simplex<\/jats:italic> and 0.0001\u2009ng\/\u00b5L for <jats:italic>P. decipiens<\/jats:italic>. The specificity test also confirmed that there was no cross-activity with the five parasite samples and the three types of anisakids plasmid DNA. This study would contribute development of a rapid detection method for anisakidosis, providing a foundation for proactive responses to food poisoning outbreaks.<\/jats:p>","DOI":"10.1371\/journal.pone.0320724","type":"journal-article","created":{"date-parts":[[2025,4,25]],"date-time":"2025-04-25T17:26:03Z","timestamp":1745601963000},"page":"e0320724","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":1,"title":["Comparative study of Taqman-based qPCR assay for the detection of Anisakis simplex and Pseudoterranova decipiens"],"prefix":"10.1371","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0009-0006-3558-5389","authenticated-orcid":true,"given":"Mi-Gyeong","family":"Kim","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Min Ji","family":"Hong","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9195-9975","authenticated-orcid":true,"given":"Doo Won","family":"Seo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hyun Mi","family":"Jung","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hyun-Ja","family":"Han","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Seung Hwan","family":"Kim","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Insun","family":"Joo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2025,4,25]]},"reference":[{"key":"pone.0320724.ref001","doi-asserted-by":"crossref","first-page":"384","DOI":"10.1016\/j.ijppaw.2019.04.007","article-title":"Anisakis Nematodes in Fish and Shellfish- from infection to allergies","volume":"9","author":"IE Aibinu","year":"2019","journal-title":"Int J Parasitol Parasites Wildl"},{"issue":"7","key":"pone.0320724.ref002","doi-asserted-by":"crossref","first-page":"2095","DOI":"10.1007\/s00436-018-5896-5","article-title":"A scanning electron microscopy study of Anisakis physeteris molecularly identified: from third stage larvae from fish to fourth stage larvae obtained in vitro","volume":"117","author":"D 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