{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T01:53:38Z","timestamp":1792115618185,"version":"4.4.0"},"reference-count":32,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2018,1,25]],"date-time":"2018-01-25T00:00:00Z","timestamp":1516838400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2018,1,25]],"date-time":"2018-01-25T00:00:00Z","timestamp":1516838400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Reverse transcription\u2013quantitative polymerase chain reaction is a valuable and reliable method for gene quantification. Target gene expression is usually quantified by normalization using reference genes (RGs), and accurate normalization is critical for producing reliable data. However, stable RGs in nasal polyps and sinonasal tissues from patients with chronic rhinosinusitis (CRS) have not been well investigated. Here, we used a two-stage study design to identify stable RGs. We assessed the stability of 15 commonly used candidate RGs using five programs\u2014geNorm, NormFinder, BestKeeper, \u0394CT, and RefFinder. Ribosomal protein lateral stalk subunit P1 (<jats:italic>RPLP1<\/jats:italic>) and ribosomal protein lateral stalk subunit P0 (<jats:italic>RPLP0<\/jats:italic>) were the two most stable RGs in the first stage of the study, and these results were validated in the second stage. The commonly used RGs \u03b2-actin (<jats:italic>ACTB<\/jats:italic>) and glyceraldehyde 3-phosphate dehydrogenase (<jats:italic>GAPDH<\/jats:italic>) were unstable according to all of the algorithms used. The findings were further validated via relative quantification of <jats:italic>IL-5, CCL11, IFN-\u03b3<\/jats:italic>, and <jats:italic>IL-17A<\/jats:italic> using the stable and unstable RGs. The relative expression levels varied greatly according to normalization with the selected RGs. Appropriate selection of stable RGs will allow more accurate determination of target gene expression levels in patients with CRS.<\/jats:p>","DOI":"10.1038\/s41598-018-19834-9","type":"journal-article","created":{"date-parts":[[2018,1,19]],"date-time":"2018-01-19T10:45:46Z","timestamp":1516358746000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":29,"title":["Assessment of suitable reference genes for RT\u2013qPCR studies in chronic rhinosinusitis"],"prefix":"10.1038","volume":"8","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8761-9292","authenticated-orcid":false,"given":"Tsuguhisa","family":"Nakayama","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Naoko","family":"Okada","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5908-1752","authenticated-orcid":false,"given":"Mamoru","family":"Yoshikawa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daiya","family":"Asaka","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Akihito","family":"Kuboki","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hiromi","family":"Kojima","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yasuhiro","family":"Tanaka","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shin-ichi","family":"Haruna","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2018,1,25]]},"reference":[{"key":"19834_CR1","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2199-10-31","volume":"10","author":"A Ho-Pun-Cheung","year":"2009","unstructured":"Ho-Pun-Cheung, A. et al. Reverse transcription-quantitative polymerase chain reaction: Description of a RIN-based algorithm for accurate data normalization. BMC Mol Biol 10, 31 (2009).","journal-title":"BMC Mol Biol"},{"key":"19834_CR2","doi-asserted-by":"publisher","first-page":"e63","DOI":"10.1093\/nar\/gkr065","volume":"39","author":"J Vermeulen","year":"2011","unstructured":"Vermeulen, J. et al. Measurable impact of RNA quality on gene expression results from quantitative PCR. Nucleic Acids Research 39, e63\u2013e63 (2011).","journal-title":"Nucleic Acids Research"},{"key":"19834_CR3","doi-asserted-by":"publisher","first-page":"187","DOI":"10.1586\/14737159.9.2.187","volume":"9","author":"J Murphy","year":"2009","unstructured":"Murphy, J. & Bustin, S. A. Reliability of real-time reverse-transcription PCR in clinical diagnostics: gold standard or substandard? Expert Rev. Mol. Diagn. 9, 187\u2013197 (2009).","journal-title":"Expert Rev. Mol. 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Journal of Allergy and Clinical Immunology 114, 155\u2013212 (2004).","journal-title":"Journal of Allergy and Clinical Immunology"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-018-19834-9","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-018-19834-9.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-018-19834-9.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,21]],"date-time":"2022-12-21T01:23:19Z","timestamp":1671585799000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-018-19834-9"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,1,25]]},"references-count":32,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2018,12]]}},"alternative-id":["19834"],"URL":"https:\/\/doi.org\/10.1038\/s41598-018-19834-9","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,1,25]]},"assertion":[{"value":"2 June 2017","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"9 January 2018","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"25 January 2018","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare that they have no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing Interests"}}],"article-number":"1568"}}