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Peer-reviewedCC BYMolecular biology reports

Identification and validation of stable reference genes for RT-qPCR analysis in blood samples from golden Syrian hamsters under different immunological conditions.

  • Jady Duarte Nogueira1
  • Tiffany Thurow Bunde1
  • Mara Andrade Colares Maia1
  • Natasha Rodrigues de Oliveira1,2
  • Odir Antônio Dellagostin1
  • Thaís Larré Oliveira Bohn3
  1. 1Laboratório de Vacinologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, Rio Grande do Sul, Brasil.
  2. 2Instituto de Ciências Biológicas, Universidade Federal do Rio Grande, Rio Grande, Rio Grande do Sul, Brazil.
  3. 3Laboratório de Vacinologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, Rio Grande do Sul, Brasil. [email protected].
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Abstract

Background

The golden Syrian hamster is the main experimental model for leptospirosis. However, there are no validated reference genes for reverse transcription quantitative PCR (RT-qPCR) analysis of blood samples.Methods and results

The expression stability of 11 candidate reference genes was evaluated in hamster blood under distinct immunological conditions, including Leptospira interrogans infection and different immunization strategies. Stability was assessed using BestKeeper, geNorm, NormFinder, and the comparative ΔCt method, with the consensus ranking calculated using the RefFinder approach implemented in the RefSeeker R package. The most stable reference gene was ppia, followed by gapdh and ppib, whereas actb was the least stable gene in the ranking. Normalization of tnf-α expression using actb resulted in an approximately 10-fold overestimation compared with ppia, demonstrating the impact of reference gene choice on data interpretation.Conclusions

These findings support ppia as a reliable reference gene for RT-qPCR normalization in hamster blood under both infection and vaccination conditions.

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Cite

BibTeXRIS
BibTeX
@article{nogueira2026identification,
  title = {{Identification and validation of stable reference genes for RT-qPCR analysis in blood samples from golden Syrian hamsters under different immunological conditions.}},
  author = {Jady Duarte Nogueira and Tiffany Thurow Bunde and Mara Andrade Colares Maia and Natasha Rodrigues de Oliveira and Odir Antônio Dellagostin and Thaís Larré Oliveira Bohn},
  journal = {Molecular biology reports},
  year = {2026},
  month = oct,
  doi = {10.1007/s11033-026-12980-8},
  url = {https://europepmc.org/article/MED/42909732},
  note = {CC BY},
}
RIS
TY  - JOUR
TI  - Identification and validation of stable reference genes for RT-qPCR analysis in blood samples from golden Syrian hamsters under different immunological conditions.
AU  - Jady Duarte Nogueira
AU  - Tiffany Thurow Bunde
AU  - Mara Andrade Colares Maia
AU  - Natasha Rodrigues de Oliveira
AU  - Odir Antônio Dellagostin
AU  - Thaís Larré Oliveira Bohn
PY  - 2026
DA  - 2026/10/23
JO  - Molecular biology reports
DO  - 10.1007/s11033-026-12980-8
UR  - https://europepmc.org/article/MED/42909732
AB  - Background  The golden Syrian hamster is the main experimental model for leptospirosis. However, there are no validated reference genes for reverse transcription quantitative PCR (RT-qPCR) analysis of blood samples.Methods and results  The expression stability of 11 candidate reference genes was evaluated in hamster blood under distinct immunological conditions, including Leptospira interrogans infection and different immunization strategies. Stability was assessed using BestKeeper, geNorm, NormFinder, and the comparative ΔCt method, with the consensus ranking calculated using the RefFinder approach implemented in the RefSeeker R package. The most stable reference gene was ppia, followed by gapdh and ppib, whereas actb was the least stable gene in the ranking. Normalization of tnf-α expression using actb resulted in an approximately 10-fold overestimation compared with ppia, demonstrating the impact of reference gene choice on data interpretation.Conclusions  These findings support ppia as a reliable reference gene for RT-qPCR normalization in hamster blood under both infection and vaccination conditions.
SN  - 0301-4851
SN  - 1573-4978
ER  -