ArticleInternational journal of molecular sciences2015
Evaluation and Selection of Appropriate Reference Genes for Real-Time Quantitative PCR Analysis of Gene Expression in Nile Tilapia (Oreochromis niloticus) during Vaccination and Infection.
Article in International journal of molecular sciences, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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17 citing papers in PubMed, 65 citations in OpenAlex.
- Dietary β-Glucan From the Marine Diatom Chaetoceros muelleri Enhances Immune Responses and Resistance of Nile Tilapia to Tilapia Lake Virus Infection.Journal of fish diseases · 2026Article
- Temporal Influence of Transport Length on Epidermal Barrier Integrity, Mucin Transcription, and Goblet Cell Kinetics in Red Tilapia (Animals : an open access journal from MDPI · 2026Article
- Screening of Reference Genes for Quantitative Real-Time PCR Analysis in Tissues and during Testis Development, and Application to Analyze the Expression ofAnimals : an open access journal from MDPI · 2024Article
- Correlation of Streptococcus agalactiae concentration on immune system and effective dose of inactivated vaccine for Chitralada 3 strain Nile tilapia (Oreochromis niloticus) in Thailand.BMC veterinary research · 2023Article
- Comparative Study ofFrontiers in cellular and infection microbiology · 2022Article
- Predator-induced maternal effects determine adaptive antipredator behaviors via egg composition.Proceedings of the National Academy of Sciences of the United States of America · 2021Article
- Identification and Evaluation of Reference Genes for Normalization of Gene Expression in Developmental Stages, Sexes, and Tissues of Diaphania caesalis (Lepidoptera, Pyralidae).Journal of insect science (Online) · 2020Article
- Identification and Characterization of Carnitine Palmitoyltransferase 1 (Evolutionary bioinformatics online · 2020Article
- Evaluation of duplicated reference genes for quantitative real-time PCR analysis in genome unknown hexaploid oat (Plant methods · 2020Article
- QTL Mapping for Red Blotches in Malaysia Red Tilapia (Oreochromis spp.).Marine biotechnology (New York, N.Y.) · 2019Article
- Validation and Evaluation of Reference Genes for Quantitative Real-Time PCR inInternational journal of molecular sciences · 2018Article
- Stable Internal Reference Genes for Normalizing Real-Time Quantitative PCR inFrontiers in plant science · 2017Article
- Interleukin-8 holds promise to serve as a molecular adjuvant in DNA vaccination model against Streptococcus iniae infection in fish.Oncotarget · 2016Article
- Molecular characterization and expression analysis of Hsp90 in Schizothorax prenanti.Cell stress & chaperones · 2016Article
- Genome-Wide Characterization and Expression Analysis of the Germin-Like Protein Family in Rice and Arabidopsis.International journal of molecular sciences · 2016Article
- Evaluation of Housekeeping Genes for Quantitative Real-Time PCR Analysis of Bradysia odoriphaga (Diptera: Sciaridae).International journal of molecular sciences · 2016Article
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12 authors at 1 institution in 1 country.
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No grant is acknowledged in the PubMed record.
Abstract
qPCR as a powerful and attractive methodology has been widely applied to aquaculture researches for gene expression analyses. However, the suitable reference selection is critical for normalizing target genes expression in qPCR. In the present study, six commonly used endogenous controls were selected as candidate reference genes to evaluate and analyze their expression levels, stabilities and normalization to immune-related gene IgM expression during vaccination and infection in spleen of tilapia with RefFinder and GeNorm programs. The results showed that all of these candidate reference genes exhibited transcriptional variations to some extent at different periods. Among them, EF1A was the most stable reference with RefFinder, followed by 18S rRNA, ACTB, UBCE, TUBA and GAPDH respectively and the optimal number of reference genes for IgM normalization under different experiment sets was two with GeNorm. Meanwhile, combination the Cq (quantification cycle) value and the recommended comprehensive ranking of reference genes, EF1A and ACTB, the two optimal reference genes, were used together as reference genes for accurate analysis of immune-related gene expression during vaccination and infection in Nile tilapia with qPCR. Moreover, the highest IgM expression level was at two weeks post-vaccination when normalized to EF1A, 18S rRNA, ACTB, and EF1A together with ACTB compared to one week post-vaccination before normalizing, which was also consistent with the IgM antibody titers detection by ELISA.
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