{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T19:18:44Z","timestamp":1792783124545,"version":"4.5.0"},"reference-count":17,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T00:00:00Z","timestamp":1790467200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T00:00:00Z","timestamp":1790467200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"name":"National Health and Medical Research Council of Australia","award":["1147897"],"award-info":[{"award-number":["1147897"]}],"id":[{"id":"https:\/\/ror.org\/011kf5r70","id-type":"ROR","asserted-by":"crossref"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Mol Biol Rep"],"published-print":{"date-parts":[[2026,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>\n                      Reliable qPCR analysis depends on stable endogenous controls, particularly in high-throughput pre-amplification workflows where technical bias may arise. The suitability of highly abundant 18\u00a0S rRNA as a reference gene in multiplex OpenArray\n                      <jats:sup>\u00ae<\/jats:sup>\n                      platforms remains debated. This study evaluated the stability of 18\u00a0S rRNA, its impact on co-amplified targets, and comparability between archived and newly manufactured primer pools.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Plasma RNA from the Fenofibrate Intervention and Event Lowering in Diabetes (FIELD) trial was analysed using TaqMan OpenArray\n                      <jats:sup>\u00ae<\/jats:sup>\n                      following reverse transcription and pre-amplification. Paired samples (\n                      <jats:italic>n<\/jats:italic>\n                      \u2009=\u200923) were pre-amplified with primer pools with and without 18\u00a0S rRNA and compared across four target genes. Stability of 18\u00a0S rRNA was assessed in 789 samples across 17 runs. Agreement between archived and new primer pools was evaluated in 774 paired samples using Wilcoxon signed-rank and Bland\u2013Altman analyses.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      18\u00a0S rRNA showed stable detection (median Ct 14.5, IQR 13.4\u201316.3) with low inter-run variability (CV 3.9%). Inclusion of 18\u00a0S rRNA during pre-amplification did not significantly alter Ct values of co-amplified targets (median differences 0.07\u20130.52 cycles;\n                      <jats:italic>p<\/jats:italic>\n                      \u2009&gt;\u20090.05), with minimal overall bias (-0.18 Ct). Archived and newly manufactured primer pools showed a small but significant Ct shift, with lower values in the new pool (bias 0.53 Ct;\n                      <jats:italic>p<\/jats:italic>\n                      \u2009&lt;\u20090.0001), although variability between paired measurements was wide.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      18\u00a0S rRNA is a stable technical control in plasma OpenArray\n                      <jats:sup>\u00ae<\/jats:sup>\n                      workflows and did not measurably bias co-amplified targets during pre-amplification. Archived and new primer pools showed only minor systematic differences and remained broadly comparable at the group level.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1007\/s11033-026-12820-9","type":"journal-article","created":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T10:26:06Z","timestamp":1790504766000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Evaluation of 18\u00a0S rRNA as an endogenous control in OpenArray\u00ae plasma RNA qPCR workflows"],"prefix":"10.1007","volume":"53","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-1106-4118","authenticated-orcid":false,"given":"Habib","family":"Francis","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0995-4524","authenticated-orcid":false,"given":"Andrzej S.","family":"Januszewski","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abubakar","family":"Mangani","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fahmida K.","family":"Ema","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anandwardhan A.","family":"Hardikar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mugdha V.","family":"Joglekar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael L.H.","family":"Huang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3085-5627","authenticated-orcid":false,"given":"David R.","family":"Sullivan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1227-803X","authenticated-orcid":false,"given":"Ronald C.W.","family":"Ma","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sanjeev","family":"Galande","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Val","family":"Gebski","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"R John","family":"Simes","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alicia J.","family":"Jenkins","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4679-5868","authenticated-orcid":false,"given":"Mark P.","family":"Molloy","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anthony C.","family":"Keech","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,9,27]]},"reference":[{"key":"12820_CR1","doi-asserted-by":"publisher","first-page":"5","DOI":"10.1186\/s12867-015-0033-9","volume":"16","author":"V Korenkova","year":"2015","unstructured":"Korenkova V, Scott J, Novosadova V, Jindrichova M, Langerova L, Svec D, Sidova M, Sjoback R (2015) Pre-amplification in the context of high-throughput qPCR gene expression experiment. 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