河南农业大学学报2011，Vol.45Issue(3)：313-317,5.
猪传染性胃肠炎病毒SYBR-Green Ⅰ荧光定量PCR检测方法的建立
Development of SYBR-Green Ⅰ real-time quantitative PCR assay for detection of transmissible gastroenteritis virus
摘要
Abstract
To develop the SYBR Green I real-time quantitative PCR assay for detection of N gene of transmissible gastroenteritis virus ( TGE V ) . A 324 bp region of the transmissible gastroenteritis virus N gene was amplified using reverse transcriptase polymerase chain reaction (RT-PCR) assay, then subcloned to pGEM-T vector and acquired the recombinant plasmid pGEM-N, which served as template to conduct the standards curve of the SYBR Green I real-time PCR. The results showed that the SYBR Green I real-time PCR was established and could used to detect TGEV. The TGEV sensitivity analysis showed that the developed SYBR Green I real-time PCR could detect 5 x 10' copy ·μL-1. And the specificity and repeatability of this method were very good. All the results suggested that the SYBR Green I real-time PCR we established in present study could be used in clinical diagnosis and epidemiological investigation for TGEV.关键词
猪传染性胃肠炎病毒/N基因/SYBR-Green Ⅰ/荧光定量PCRKey words
transmissible gastroenteritis virus/ N gene/ SYBR-Green I/ real-time quantitative PCR assay分类
农业科技引用本文复制引用
李金磊,王亚宾,崔保安,陈丽颖,张红英,李珂珂,魏战勇..猪传染性胃肠炎病毒SYBR-Green Ⅰ荧光定量PCR检测方法的建立[J].河南农业大学学报,2011,45(3):313-317,5.基金项目
教育部博士点新教师基金项目(20104105120005) （20104105120005）
河南省教育厅自然科学基金项目(2010A230007) （2010A230007）
