中国动物传染病学报2018，Vol.26Issue(3)：28-33,6.
猪源干扰素刺激基因PPBP荧光定量PCR检测方法的建立及初步应用
DEVELOPMENT AND PRELIMINARY APPLICATION OF SYBR GREEN I REAL-TIME PCR FOR DETECTION AND QUANTITATION OF PORCINE PRO-PLATELET BASIC PROTEIN
摘要
Abstract
The study was to develop a real-time assay for detecting swine pro-platelet basic protein. According to PPBP gene sequence, specific primer was designed by primer design software Primer Premier5.0 The resulting fragments were cloned into the vector to construct recombinant plasmids of p3XFLAG-CMV-7.1-PPBP. The recombinant plasmid was used as standard products to establish standard curve of porcine PPBP and detect repeatability, sensitivity and specificity. The results show a precise linear relationship with a correlation coefficient R2＞0.99. The amplification efficiency is 102%. Dissolve curve appears one peak. The variation coefficient is less than 0.5% within and between assays. The SYBR GreenI real-time PCR assay was used to detect porcine PPBP from mock pigs and pigs infected with PRRSV. PPBP was found in this samples and was significant different between the infected and mock group.This SYBR Green I real-time PCR assay developed in this study has highly sensitivity, specificity, stability and repeatability, which could be a tool for detecting relationship between pig infectious diseases and porcine PPBP.关键词
促血小板碱性蛋白/SYBR GreenI/实时荧光定量PCRKey words
Pro-platelet basic protein/SYBR GreenI/real-time PCR分类
农业科技引用本文复制引用
王飞飞,马志永,JUNG Yong-Sam,刘珂,郇贝丽,魏建超,邵东华,李蓓蓓,邱亚峰,石元元,钱莺娟..猪源干扰素刺激基因PPBP荧光定量PCR检测方法的建立及初步应用[J].中国动物传染病学报,2018,26(3):28-33,6.基金项目
973项目(2014CB542702) （2014CB542702）
