水禽圆环病毒双重PCR检测方法的建立  

Establishment of Duplex PCR for Detection of Duck Circovirus and Goose Circovirus

作　　者：曹慧慧[1] 孙文超[1] 郑敏[2] 苏姣秀[2] 梁晟[2] 张红云 韦显凯[2] 刘棋[2] 

机构地区：[1]广西大学动物科学技术学院,广西南宁530001 [2]广西动物疫病预防控制中心,广西南宁530001 [3]玉林市动物疫病预防控制中心,广西玉林537000

出　　处：《中国家禽》2015年第22期20-23,共4页China Poultry

基　　金：广西自然科学基金项目(2012GXNSFAA053073);广西水产畜牧兽医局科技计划[桂渔牧科(1204936)]

摘　　要：为建立能同时检测并区分鸭圆环病毒（Duck circovirus，DuCV）和鹅圆环病毒（G00se circovims，GOCV）的方法，根据DuCV和GoCV基因组的保守区域设计两对特异性引物，预期特异性扩增DuCV和GoCV片段大小分别为192bp、556bp。经过反应条件的优化，建立了DuCV和GoCV的双重PCR检测方法。特异性检验表明该方法无法扩增其他水禽病原；DuCV和GoCV的最低检出限分别为10^4拷贝／μL和10^5拷贝／μL。临床样品检测结果显示，DuCV阳性率为8．01％，GoCV阳性率为1．6％，与常规PCR方法完全一致。结果表明，建立的双重PCR方法可以用于水禽圆环病毒检测和流行病学调查。To develop a duplex PCR assay for detection and differentiation of duck circovirus （DuCV） and goose circovirus （GoCV）, two pairs of specific primers were designed according to the sequences of DuCV and GoCV, of which one specifically amplified a 192 bp fragment of DuCV,the other amplified a 556 bp fragment of GoCV. And the reaction conditions were optimized. Specificity test showed no amplification were observed when other common pathogens originated from waterfowl served as template. The detection limits of assay were 104 copies/μL for DuCV DNA and 10^5 copies/μL for GoCV DNA. Furthemore, the established assay was used to detect clinical samples. The results showed that the positive rates of DuCV and GoCV were 8.01% and 1.6% respectively, which were fully corresponding with those of the conventional PCR assay. These results suggested that the duplex PCR assay developed by this study was suitable for clinical detection and epidemiology surveillance of DuCV and GoCV.

关 键 词：鸭圆环病毒 鹅圆环病毒 双重PCR 

分 类 号：S855.3[农业科学—临床兽医学]