title: Real-time qPCR identifies suitable reference genes for Borna disease virus-infected rat cortical neurons.
abstract: 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.
authors.affiliation: Department of Neurology, Yongchuan Hospital, Chongqing Medical University, Chongqing 402460, China. zhlbright@gmail.com.
authors.forename: Lujun
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Chongqing Key Laboratory of Neurobiology, Chongqing 400016, China. 18983145528@163.com.
authors.forename: Siwen
authors.initials: S
authors.lastname: Liu
authors.affiliation: Chongqing Key Laboratory of Neurobiology, Chongqing 400016, China. zhlbright@gmail.com.
authors.forename: Liang
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Chongqing Key Laboratory of Neurobiology, Chongqing 400016, China. yhm0709@126.com.
authors.forename: Hongmin
authors.initials: H
authors.lastname: You
authors.affiliation: Department of Rehabilitation, the Second Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China. uang@live.com.
authors.forename: Rongzhong
authors.initials: R
authors.lastname: Huang
authors.affiliation: Chongqing Key Laboratory of Neurobiology, Chongqing 400016, China. tilamisu789456123@126.com.
authors.forename: Lin
authors.initials: L
authors.lastname: Sun
authors.affiliation: Chongqing Key Laboratory of Neurobiology, Chongqing 400016, China. hepeng000@sina.com.
authors.forename: Peng
authors.initials: P
authors.lastname: He
authors.affiliation: Chongqing Key Laboratory of Neurobiology, Chongqing 400016, China. iconsig@sina.com.
authors.forename: Shigang
authors.initials: S
authors.lastname: Chen
authors.affiliation: Chongqing Key Laboratory of Neurobiology, Chongqing 400016, China. ASDFG43215@126.com.
authors.forename: Hong
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Department of Neurology, Yongchuan Hospital, Chongqing Medical University, Chongqing 402460, China. xiepeng@cqmu.edu.cn.
authors.forename: Peng
authors.initials: P
authors.lastname: Xie
mesh_terms: D000818:Animals; D001890:Borna Disease / Q000235:genetics* / Q000821:virology*; D001891:Borna disease virus / Q000502:physiology*; D019208:Brain-Derived Neurotrophic Factor / Q000235:genetics / Q000378:metabolism; D002540:Cerebral Cortex / Q000473:pathology*; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation; D009474:Neurons / Q000378:metabolism / Q000473:pathology / Q000821:virology*; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000592:standards*; D012015:Reference Standards; D015203:Reproducibility of Results; D012984:Software
chemical_list: D019208:Brain-Derived Neurotrophic Factor