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Screening of Stable Reference Genes in <i>Solea senegalensis</i> Juveniles for Nervous Necrosis Virus (NNV) Vaccination Assays.

Overview

Authors: Lucía Vázquez-Salgado1, Carmen López-Vázquez1, Isabel Bandín1, Sandra Souto1
ORCID iDs: Lucía Vázquez-Salgado, Carmen López-Vázquez, Isabel Bandín, Sandra Souto
  1. Aquatic One Health Research Center (ARCUS), Universidade de Santiago de Compostela (USC), 15782 Santiago de Compostela, Spain; (C.L.-V.); (I.B.)
Institutions: Universidade de Santiago de Compostela (Spain)
Journal: Veterinary sciences, volume 13, issue 8, article 790
Dates: received 23 June 2026; accepted 5 August 2026; published online 8 August 2026
Type: Research article · Language: English
License: CC BY
Identifiers: DOI 10.3390/vetsci13080790 · PMID 42655810 · PMCID PMC13517329 · OpenAlex W7202069598
Open access: gold, a free copy (OpenAlex)
Status: data only
Categories: other (modality), cellular / molecular (subfield)
Methods: Statistics
Keywords: reference gene, gene expression, normalisation, rps4, ubq, RT-qPCR
Topic: Aquaculture disease management and microbiota (Immunology, Immunology and Microbiology), according to OpenAlex
Funding: European Union (101084204); Ministerio de Ciencia, Innovación y Universidades (JDC2023-051769-I, RYC2023-045820-I)
Citations: not cited yet (Europe PMC); 47 references in the paper

Abstract

Quantifying the expression of immune-related genes is essential for evaluating vaccine efficacy. However, obtaining reliable RT-qPCR results requires the use of stable reference genes. In this study, the expression stability of eight candidate reference genes (actb1, actb2, eef1a, gapdh2, hprt1, ubq, 18S, and rps4) was evaluated in brain, kidney, and gut tissues from naïve and virus-infected juvenile sole. Gene stability was assessed through two statistical algorithms (geNorm and NormFinder) and the integrative tool RefFinder, and the results were integrated through RankAggreg for a robust consensus ranking. Among the tested genes, rps4 and ubq showed the highest stability across tissues and experimental conditions and were sufficient for accurate normalisation. The suitability of these genes was validated in vaccinated sole for normalisation of the expression of proinflammatory cytokines (il1β, il6, tnfα), supporting their recommendation as optimal reference genes. Conversely, actb1 and hprt1 were the least stable genes and compromised the reliable quantification of proinflammatory cytokines in vaccinated fish.

Reproduced under the paper's license (CC BY), from the paper cited above.

Code

The paper links to its data, not to its authors' code: see the Data section.

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Data

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Data Availability Statement

The original data presented in the study are openly available in Zenodo at 10.5281/zenodo.17367550.

Reproduced under the paper's license (CC BY), from the paper cited above.

Versions

The history of this record: each version stored by the harvester or made by a correction of its authors or of the maintainers of its code, and what changed in its facts. The texts of the paper (its abstract, its availability statements) are not part of it; versions that changed only those are not listed.

Version 1, 27 September 2026: the first record

Recorded: type, language, journal, volume, issue, pages, dates, 4 authors, 6 keywords, 2 funders, 44 references.

Cite

This paper

Vázquez-Salgado, L., López-Vázquez, C., Bandín, I., & Souto, S. (2026). Screening of Stable Reference Genes in <i>Solea senegalensis</i> Juveniles for Nervous Necrosis Virus (NNV) Vaccination Assays. Veterinary sciences, 13(8), 790. https://doi.org/10.3390/vetsci13080790

BibTeX

@article{vazquezsalgado2026screening,
author = {Vázquez-Salgado, Lucía and López-Vázquez, Carmen and Bandín, Isabel and Souto, Sandra},
title = {{Screening of Stable Reference Genes in \<i\>Solea senegalensis\</i\> Juveniles for Nervous Necrosis Virus (NNV) Vaccination Assays}},
journal = {Veterinary sciences},
year = {2026},
month = aug,
volume = {13},
number = {8},
pages = {790},
publisher = {Multidisciplinary Digital Publishing Institute (MDPI)},
issn = {2306-7381},
doi = {10.3390/vetsci13080790},
url = {https://doi.org/10.3390/vetsci13080790},
pmid = {42655810},
pmcid = {PMC13517329}
}

RIS

TY - JOUR
AU - Vázquez-Salgado, Lucía
AU - López-Vázquez, Carmen
AU - Bandín, Isabel
AU - Souto, Sandra
TI - Screening of Stable Reference Genes in <i>Solea senegalensis</i> Juveniles for Nervous Necrosis Virus (NNV) Vaccination Assays
T2 - Veterinary sciences
J2 - Vet Sci
PY - 2026
DA - 2026/08/08
VL - 13
IS - 8
SP - 790
SN - 2306-7381
PB - Multidisciplinary Digital Publishing Institute (MDPI)
DO - 10.3390/vetsci13080790
UR - https://doi.org/10.3390/vetsci13080790
LA - en
ER -

CSL-JSON

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"id": "10.3390/vetsci13080790",
"type": "article-journal",
"title": "Screening of Stable Reference Genes in <i>Solea senegalensis</i> Juveniles for Nervous Necrosis Virus (NNV) Vaccination Assays",
"container-title": "Veterinary sciences",
"author": [
{
"family": "Vázquez-Salgado",
"given": "Lucía"
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"given": "Carmen"
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"container-title-short": "Vet Sci",
"volume": "13",
"issue": "8",
"page": "790",
"DOI": "10.3390/vetsci13080790",
"PMID": "42655810",
"PMCID": "PMC13517329",
"ISSN": "2306-7381",
"publisher": "Multidisciplinary Digital Publishing Institute (MDPI)",
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"language": "en",
"issued": {
"date-parts": [
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}

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