Real-Time qPCR Identifies Suitable Reference Genes for Borna Disease Virus-Infected Rat Cortical Neurons

International Journal of Molecular Sciences

Lujun Zhang²  Siwen Liu¹  Liang Zhang¹  Hongmin You¹  Rongzhong Huang³  Lin Sun¹  Peng He¹  Shigang Chen¹  Hong Zhang¹  Peng Xie² 

[1] Chongqing Key Laboratory of Neurobiology, Chongqing 400016, China; E-Mails:;Department of Neurology, Yongchuan Hospital, Chongqing Medical University, Chongqing 402460, China; E-Mail:;Department of Rehabilitation, the Second Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China; E-Mail:

关键词: Borna disease virus;    BDV;    reference gene;    RT-qPCR;    cortical neuron;   

DOI  :  10.3390/ijms151221825

来源: mdpi

【 摘 要 】

Quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR) is the most commonly-used technique to identify gene expression profiles. The selection of stably expressed reference genes is a prerequisite to properly evaluating gene expression. Here, the suitability of commonly-used reference genes in normalizing RT-qPCR assays of mRNA expression in cultured rat cortical neurons infected with Borna disease virus (BDV) was assessed. The expressions of eight commonly-used reference genes were comparatively analyzed in BDV-infected rat cortical neurons and non-infected control neurons mainly across 9 and 12 days post-infection. These reference genes were validated by RT-qPCR and separately ranked by four statistical algorithms: geNorm, NormFinder, BestKeeper and the comparative delta-Ct method. Then, the RankAggreg package was used to construct consensus rankings. ARBP was found to be the most stable internal control gene at Day 9, and ACTB at Day 12. As the assessment of the validity of the selected reference genes confirms the suitability of applying a combination of the two most stable references genes, combining the two most stable genes for normalization of RT-qPCR studies in BDV-infected rat cortical neurons is recommended at each time point. This study can contribute to improving BDV research by providing the means by which to obtain more reliable and accurate gene expression measurements.

【 授权许可】

CC BY   
© 2014 by the authors; licensee MDPI, Basel, Switzerland.