中国畜牧兽医2016，Vol.43Issue(10)：2534-2540,7.DOI:10.16431/j.cnki.1671-7236.2016.10.004
Nsp9基因实时荧光定量PCR检测方法的建立及其在感染细胞过程中表达量的变化
Establishment of the Quantitative Real-time PCR Method for PRRSV Nsp9 Gene Rapid Detection and Its Expression in PRRSV Infected Cells
摘要
Abstract
The study was conducted to establish a quantitative Real-time PCR method for PRRSV Nsp 9 gene rapid detection and study its expression in PRRSV infected cells.A pair of specific primers targeted to Nsp9 of PRRSV was designed and a Real-time PCR method based on SYBR Green Ⅰ fluorescent was developed for the quantization of PRRSV.The melting curve analysis u-sing SYBR Green Ⅰ dye showed one specific peak,and no primer dimers peak was observed.No amplification was detected from HEV,SIV and PRV samples by this method.There was good re-producibility and low variation coefficient.The quantitative Real-time PCR method developed in this study would be useful for rapid laboratory diagnosis and epidemiology investigation of PRRSV.During the process of virus infecting cells,the expression level of Nsp9 increased gradu-ally,and it got the highest at 36 h.This study laid the theoretical basis to explore the law of virus replication and clinical vaccine production.关键词
猪繁殖与呼吸综合征病毒/SYBRGreenⅠ/实时荧光定量PCR/Nsp9基因Key words
PRRSV/SYBR Green Ⅰ/quantitative Real-time PCR/Nsp 9 gene分类
农业科技引用本文复制引用
赵孟孟,冯松林,王文佳,邢星,冯嘉萍,张桂红..Nsp9基因实时荧光定量PCR检测方法的建立及其在感染细胞过程中表达量的变化[J].中国畜牧兽医,2016,43(10):2534-2540,7.基金项目
国家自然科学基金猪繁殖与呼吸综合征病毒非结构蛋白 Nsp9功能研究项目(31272564)；公益性行业(农业)科研专项经费(201203039)；国家生猪现代农业产业技术体系(CARS-36)项目；国家重点研发计划课题(2016YFD0500707) （31272564）
