检测2种基因型鸭圆环病毒双重PCR方法的建立与应用  

Development and application of duplex PCR for the detection of two genotypes of duck circovirus

作　　者：李志国[1] 王鑫[1] 张瑞华[1] 陈君豪[1] 夏琳琳[1] 林少莉 谢之景[1] 姜世金[1] 

机构地区：[1]山东农业大学动物医学院/山东省动物生物工程与疾病防治重点实验室,山东泰安271018

出　　处：《中国兽医学报》2015年第7期1060-1063,1068,共5页Chinese Journal of Veterinary Science

基　　金：山东省自然科学基金资助项目(ZR2013CM009);山东省现代农业产业技术体系家禽创新团队资助项目(SDAIT-13-011-15)

摘　　要：为了研究2种基因型鸭圆环病毒(Duck circovirus,DuCV)在我国鸭群中的混合感染情况,根据鸭圆环Cap蛋白基因序列设计了4条特异性引物,建立了可以同时检测2种基因型DuCV的双重PCR检测方法。结果显示,该方法特异性强,灵敏度高,最低可检测到1拷贝的DuCV核酸。使用建立的双重PCR方法对收集的山东省6个鸭群150只病死雏鸭的组织DNA进行检测,结果显示有50.0%(3/6)的鸭群检出DuCV-1和DuCV-2的混合感染,33.33%(2/6)的鸭群检出DuCV-2的单独感染。19只雏鸭被检出感染DuCV,其中2.67%(4/150)的雏鸭检出DuCV-1和DuCV-2的混合感染,10.0%(15/150)的雏鸭只感染DuCV-1或DuCV-2。结果表明,山东省鸭群中存在较为严重的DuCV-1和DuCV-2混合感染的现象。In order to study the mixed infection of duck circovirus（DuCV）two genotypes in China,according to the capsid protein gene sequences,four specific primers were designed for development of a duplex RT-PCR assay to identify DuCV-1and DuCV-2with mixed infection.The method was shown to be high specificity and sensitivity.The minimum detection limit of the method was determined to be one copy viral DNA of DuCV-1and DuCV-2,respectively.Using the method,different tissue DNA samples of 150 dead ducklings from 6flocks in Shandong province of China were detected,50.0%（3/6）of flocks were identified as mixed infection of DuCV-1,DuCV-2,and 33.33%（2/6）of flocks were DuCV-2single infection.Among them,2.67%（4/150）of ducklings were detected as mixed infection of DuCV-1and DuCV-2,and 10.0%（15/150）of ducklings were DuCV-1or DuCV-2single infection.These results indicated that there are a certain extent of DuCV-1DuCV-2mixed infection in duck flocks in Shandong province.

关 键 词：鸭圆环病毒 双重PCR 基因型 检测 

分 类 号：S852.65[农业科学—基础兽医学]