作 者：
郭翠丽;张春红;郭鹏举;张建峰;沈海燕;潘家强
单 位：
华南农业大学;广东省农业科学院动物卫生研究所
关键词：
鸡传染性贫血病病毒(CAV);TaqMan实时荧光定量PCR;探针
摘 要：
根据鸡传染性贫血病病毒(chicken infectious anemia virus,CAV)基因组保守区域设计1对特异性引物和探针,通过优化反应条件,建立了一种快速检测CAV的TaqMan实时荧光定量PCR方法,同时验证其特异性、灵敏性和重复性。本试验建立的TaqMan实时荧光定量PCR方法灵敏度可达1.8×101拷贝/μL,远高于常规PCR方法;并与禽类其他病毒性疾病无交叉反应,具有高特异性。用分离的病毒人工感染1日龄SPF雏鸡,14日龄剖检,对感染鸡体内各器官的病毒分布及载量进行检测,结果表明,在脑、心脏、肝脏、脾脏、肺脏、肾脏、胸腺、法氏囊和血清中均可检测到病毒,肝脏、胸腺病毒载量明显高于其他组织。本研究建立的TaqMan实时荧光定量PCR方法特异性强、灵敏度高、重复性好,可同时检测大量临床样品,适用于CAV的诊断与流行病学分析。
译 名：
Development of TaqMan Fluorescence Quantitative PCR Assay for Detection of Chicken Infectious Anemia Virus
作 者：
GUO Cui-li;ZHANG Chun-hong;GUO Peng-ju;ZHANG Jian-feng;SHEN Hai-yan;PAN Jia-qiang;South China Agricultural University;Institute of Animal Health,Guangdong Academy of Agricultural Sciences;
关键词：
chicken infectious anemia virus;;TaqMan Real-time PCR;;probe
摘 要：
A fluorescence quantitative PCR assay was developed using primers and probes designed according to a high conservative sequence of chicken infectious anemia virus(CAV).Detection of viral DNA copy number was determined by extrapolation from a CAV plasmid-based standard curve.The results showed that the estimated average viral copy number could detect1.8×101 copies/μL of plasmid DNA with high reproducibility(CAV of the intra or inter)assay was below 2%.Furthermore,1day-old SPF chicken were inoculated with CAV and the virus loads were detected in multiple organs.The results showed that the virus could be detected in the brain,heart,liver,spleen,lung,kidney,thymus,bursa and serum and the virus loads in these organs showed no significant difference.No cross-reaction was detected with other avian virus.
