{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,8]],"date-time":"2026-05-08T17:15:36Z","timestamp":1778260536163,"version":"3.51.4"},"reference-count":70,"publisher":"SAGE Publications","issue":"1","license":[{"start":{"date-parts":[[2023,11,2]],"date-time":"2023-11-02T00:00:00Z","timestamp":1698883200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.sagepub.com\/page\/policies\/text-and-data-mining-license"}],"funder":[{"DOI":"10.13039\/501100003141","name":"consejo nacional de ciencia y tecnolog\u00eda","doi-asserted-by":"publisher","award":["Becas CONACYT-Regional Noroeste 2020"],"award-info":[{"award-number":["Becas CONACYT-Regional Noroeste 2020"]}],"id":[{"id":"10.13039\/501100003141","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.sagepub.com"],"crossmark-restriction":true},"short-container-title":["J VET Diagn Invest"],"published-print":{"date-parts":[[2024,1]]},"abstract":"<jats:p>\n                    Normalization, the process of controlling for normal variation in sampling and testing, can be achieved in real-time PCR assays by converting sample quantification cycles (Cqs) to \u201cefficiency standardized Cqs\u201d (ECqs). We calculated ECqs as E\n                    <jats:sup>\u2212\u0394Cq<\/jats:sup>\n                    , where E is amplification efficiency and \u0394Cq is the difference between sample and reference standard Cqs. To apply this approach to a commercial porcine reproductive and respiratory syndrome virus (PRRSV) RT-qPCR assay, we created reference standards by rehydrating and then diluting (1\u2009\u00d7\u200910\n                    <jats:sup>\u22124<\/jats:sup>\n                    ) a PRRSV modified-live vaccine (PRRS MLV; Ingelvac) with serum or oral fluid (OF) to match the sample matrix to be tested. Sample ECqs were calculated using the mean E and reference standard Cq calculated from the 4 reference standards on each plate. Serum ( n\u2009=\u2009132) and OF ( n\u2009=\u2009130) samples were collected from each of 12 pigs vaccinated with a PRRSV MLV from \u22127 to 42\u2009d post-vaccination, tested, and sample Cqs converted to ECqs. Mean plate Es were 1.75\u20132.6 for serum and 1.7\u20132.3 for OF. Mean plate reference standard Cqs were 29.1\u201331.3 for serum and 29.2\u201331.5 for OFs. Receiver operating characteristic analysis calculated the area under the curve for serum and OF sample ECqs as 0.999 (95% CI: 0.997, 1.000) and 0.947 (0.890, 1.000), respectively. For serum, diagnostic sensitivity and specificity of the commercial PRRSV RT-qPCR assay were estimated as 97.9% and 100% at an ECq cutoff \u2265\u20090.20, and for OF, 82.6% and 100%, respectively, at an ECq cutoff \u2265\u20090.45.\n                  <\/jats:p>","DOI":"10.1177\/10406387231206080","type":"journal-article","created":{"date-parts":[[2023,11,3]],"date-time":"2023-11-03T02:18:10Z","timestamp":1698977890000},"page":"78-85","update-policy":"https:\/\/doi.org\/10.1177\/sage-journals-update-policy","source":"Crossref","is-referenced-by-count":5,"title":["Normalizing real-time PCR results in routine testing"],"prefix":"10.1177","volume":"36","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4094-6813","authenticated-orcid":false,"given":"Betsy","family":"Armenta-Leyva","sequence":"first","affiliation":[{"name":"Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Lloyd Veterinary Medical Center, Iowa State University, Ames, IA, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2557-1817","authenticated-orcid":false,"given":"Berenice","family":"Mungu\u00eda-Ram\u00edrez","sequence":"additional","affiliation":[{"name":"Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Lloyd Veterinary Medical Center, Iowa State University, Ames, IA, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3732-9147","authenticated-orcid":false,"given":"Ting-Yu","family":"Cheng","sequence":"additional","affiliation":[{"name":"Department of Veterinary Preventive Medicine, College of Veterinary Medicine, the Ohio State University, Columbus, OH, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6994-1210","authenticated-orcid":false,"given":"Fangshu","family":"Ye","sequence":"additional","affiliation":[{"name":"Department of Statistics, College of Liberal Arts and Sciences, Iowa State University, Ames, IA, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alexandra","family":"Henao-D\u00edaz","sequence":"additional","affiliation":[{"name":"Pig Improvement M\u00e9xico, Santiago de Quer\u00e9taro, Quer\u00e9taro, M\u00e9xico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4407-7996","authenticated-orcid":false,"given":"Luis G.","family":"Gim\u00e9nez-Lirola","sequence":"additional","affiliation":[{"name":"Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Lloyd Veterinary Medical Center, Iowa State University, Ames, IA, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8107-2422","authenticated-orcid":false,"given":"Jeffrey","family":"Zimmerman","sequence":"additional","affiliation":[{"name":"Department of Veterinary Diagnostic and Production Animal 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