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However, there are no validated reference genes for reverse transcription quantitative PCR (RT-qPCR) analysis of blood samples.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods and Results<\/jats:title>\n                    <jats:p>\n                      The expression stability of 11 candidate reference genes was evaluated in hamster blood under distinct immunological conditions, including\n                      <jats:italic>Leptospira interrogans<\/jats:italic>\n                      infection and different immunization strategies. Stability was assessed using BestKeeper, geNorm, NormFinder, and the comparative \u0394Ct method, with the consensus ranking calculated using the RefFinder approach implemented in the RefSeeker R package. The most stable reference gene was\n                      <jats:italic>ppia<\/jats:italic>\n                      , followed by\n                      <jats:italic>gapdh<\/jats:italic>\n                      and\n                      <jats:italic>ppib<\/jats:italic>\n                      , whereas\n                      <jats:italic>actb<\/jats:italic>\n                      was the least stable gene in the ranking. Normalization of\n                      <jats:italic>tnf-\u03b1<\/jats:italic>\n                      expression using\n                      <jats:italic>actb<\/jats:italic>\n                      resulted in an approximately 10-fold overestimation compared with\n                      <jats:italic>ppia<\/jats:italic>\n                      , demonstrating the impact of reference gene choice on data interpretation.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>\n                      These findings support\n                      <jats:italic>ppia<\/jats:italic>\n                      as a reliable reference gene for RT-qPCR normalization in hamster blood under both infection and vaccination conditions.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1007\/s11033-026-12980-8","type":"journal-article","created":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T10:10:26Z","timestamp":1792750226000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Identification and validation of stable reference genes for RT-qPCR analysis in blood samples from golden Syrian hamsters under different immunological conditions"],"prefix":"10.1007","volume":"53","author":[{"given":"Jady Duarte","family":"Nogueira","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tiffany Thurow","family":"Bunde","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mara Andrade Colares","family":"Maia","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Natasha Rodrigues","family":"de Oliveira","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Odir Ant\u00f4nio","family":"Dellagostin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5330-1462","authenticated-orcid":false,"given":"Tha\u00eds Larr\u00e9 Oliveira","family":"Bohn","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,10,23]]},"reference":[{"key":"12980_CR1","doi-asserted-by":"publisher","first-page":"835","DOI":"10.1007\/s10096-019-03797-4","volume":"39","author":"KB Karpagam","year":"2020","unstructured":"Karpagam KB, Ganesh B (2020) Leptospirosis: a neglected tropical zoonotic infection of public health importance\u2014an updated review. 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The founding sponsors had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, and in the decision to publish the results.","order":1,"name":"Ethics","label":"Conflict of interest","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The animal experimental procedures described herein were approved by the Animal Ethics Committee of the Federal University of Pelotas (UFPel), under protocol number 022906\/2023-38. All animals included in this study were maintained and handled in accordance with ethical guidelines for the use of animals in research, as established by the National Council for the Control of Animal Experimentation (CONCEA).","order":2,"name":"Ethics","label":"Ethics approval","group":{"name":"EthicsHeading","label":"Declarations"}}],"article-number":"1791"}}