Mehrbod Parvaneh, Harun Mohammad Syamsul Reza, Shuid Ahmad Naqib, Omar Abdul Rahman
Institute of Bioscience, Faculty of Veterinary Medicine, Universiti Putra Malaysia, UPM Serdang, 43400, Selangor, Malaysia.
Methods Mol Biol. 2015;1282:241-50. doi: 10.1007/978-1-4939-2438-7_20.
Feline infectious peritonitis (FIP) is a lethal systemic disease caused by FIP virus (FIPV). There are no effective vaccines or treatment available, and the virus virulence determinants and pathogenesis are not fully understood. Here, we describe the sequencing of RNA extracted from Crandell Rees Feline Kidney (CRFK) cells infected with FIPV using the Illumina next-generation sequencing approach. Bioinformatics analysis, based on Felis catus 2X annotated shotgun reference genome, using CLC bio Genome Workbench is used to map both control and infected cells. Kal's Z test statistical analysis is used to analyze the differentially expressed genes from the infected CRFK cells. In addition, RT-qPCR analysis is used for further transcriptional profiling of selected genes in infected CRFK cells and Peripheral Blood Mononuclear Cells (PBMCs) from healthy and FIP-diagnosed cats.
猫传染性腹膜炎(FIP)是一种由猫传染性腹膜炎病毒(FIPV)引起的致命性全身性疾病。目前尚无有效的疫苗或治疗方法,且病毒毒力决定因素和发病机制尚未完全明确。在此,我们描述了使用Illumina下一代测序方法对感染FIPV的克兰德尔·里斯猫肾(CRFK)细胞中提取的RNA进行测序的过程。基于家猫2X注释鸟枪法参考基因组,使用CLC生物基因组工作台进行生物信息学分析,以对对照细胞和感染细胞进行定位。卡尔Z检验统计分析用于分析感染的CRFK细胞中差异表达的基因。此外,逆转录定量聚合酶链反应(RT-qPCR)分析用于对感染的CRFK细胞以及来自健康猫和FIP确诊猫的外周血单个核细胞(PBMC)中选定基因进行进一步的转录谱分析。