动物医学进展2016，Vol.37Issue(4)：12-17,6.
鸡三种天然免疫相关受体和配体荧光定量PCR 检测方法的建立及应用
Development and Application of Quantitative PCR Assay for Detection of Three Innate Immunity-related Cellular Receptors and Ligands in Chickens
摘要
Abstract
The main purpose of this study is to establish quantitative PCR (qPCR)for the detection of three pattern recognition receptors (chicken Toll-like receptor 3,Toll-like receptor 7,and melanoma differentia-tion-associated protein 5)and their respective ligands (TRIF,MyD88 and MAVS)in chickens.We designed and screened PCR primers for the 6 genes.Specific PCR products of the genes were utilized to construct re-combinant plasmids,which served as template standards to establish standard curves for the assays.The as-says developed in this study were sensitive,specific and could stably detect related genes of wide concentra-tion ranges.Further,the assays were applied in the analysis of related gene expression in specific-pathogen-free (SPF)chickens after immunization with oil-emulsion vaccine against Newcastle disease virus.The up-regulations of expression of all 6 genes were peeked at six hours post vaccination,and at the same time,the mRNA level of IFN-βincreased to 252 times.This indicated the efficacy of innate immunity signaling after vaccination.In conclusion,the qPCR developed in this study may be applied in viral pathogenesis research, and novel vaccine,immnopotentiator and anti-viral drug development.关键词
鸡/天然免疫/受体与配体/检测/实时荧光定量 PCRKey words
chicken/innate immunity/receptor and ligand/detection/quantitative PCR分类
农业科技引用本文复制引用
黄艳艳,高丹丹,许传田,朱曼玲,杨少华,黄庆华,张琳,张秀美,吴家强..鸡三种天然免疫相关受体和配体荧光定量PCR 检测方法的建立及应用[J].动物医学进展,2016,37(4):12-17,6.基金项目
山东省畜禽疫病防治与繁育重点实验室主任基金项目（2015SYSZR04）；十二五国家科技支撑计划项目（2015BAD12B03）；公益性行业（农业）科研专项（201303033）；现代农业产业技术体系建设专项资金 （）
