中国动物传染病学报2017，Vol.25Issue(2)：56-60,5.
禽呼肠孤病毒TaqMan实时荧光RT-PCR方法的建立及病毒组织分布比较研究
DEVELOPMENT AND APPLICATION OF ATAQMAN RT-PCR ASSAY FOR DETECTION OF AVIAN REOVIRUSES IN TISSUES
摘要
Abstract
A Real-time PCR method for detection of Avian reoviruses was developed using a pair of primers and specific Taqman probe that were designed according to the σC genes of Avian reovirus isolates and reference strains from GenBank. The assay was optimized to produce linearity from 1×101 copies /μL to 1×1010 copies /μL in standard curve. The detection limit of the assay was 10 copies of viral RNA, which showed higher sensitivity than the currently available PCR methods. There was no cross-reaction with other common Avian viruses. Furthermore, 1-day-old SPF chickens were inoculated with the Avian reovirus isolate MS01 and reference strain S1133. The viral loads in multiple organs were detected using the PCR method developed here. The results showed that the mortality of MS01 and S1133 was 80% and 46.7%, respectively. The viruses replicated abundantly in livers, spleens and lungs. However, viral loads of the field isolate MS01 were significantly higher than those of the reference S1133. Besides, MS01 replicated well in kidneys while S1133 did poorly. The RT-PCR method developed in the present study could be used for diagnosis and molecular pathogenesis mechanism of highly pathogenic Avian reovirus.关键词
禽呼肠孤病毒/σC基因/TaqmanRT-PCR/组织分布/致病性Key words
Avian reovirus/σC gene/Taqman RT-PCR/tissue distribution/pathogenicity分类
农业科技引用本文复制引用
钟丽,孙美玉,高立,刘永振,李凯,祁小乐,高玉龙,王笑梅..禽呼肠孤病毒TaqMan实时荧光RT-PCR方法的建立及病毒组织分布比较研究[J].中国动物传染病学报,2017,25(2):56-60,5.基金项目
现代农业产业技术体系建设专项基金(CARS-42-G07) （CARS-42-G07）
