猪圆环病毒2型TaqMan实时荧光定量PCR检测方法的建立  

Establishment of TaqMan Real-time Quantitative PCR Method for Detection of PCV2

作　　者：曾思遥[1,2] 张淑琼[1] 余绍华[1] 唐志玲[1] 陈瑞爱[2] 罗满林[1,2] 

机构地区：[1]华南农业大学,广东广州510642 [2]广东大华农动物保健品股份有限公司研发中心,广东云浮527300

出　　处：《中国畜牧兽医》2012年第6期41-46,共6页China Animal Husbandry & Veterinary Medicine

基　　金：省部产学研结合重大项目(2011A090200117);广东大华农科研基金(033A201005-3)

摘　　要：本研究以猪圆环病毒2型(PCV2)核衣壳蛋白ORF2基因为目的基因,设计合成特异性引物和探针。PCR扩增得到目的基因,并克隆到pMD18-T载体,筛选得到标准阳性质粒。通过对荧光定量PCR反应条件的优化,建立了PCV2的TaqMan实时荧光定量PCR检测方法。试验结果表明,该方法特异性强,对猪伪狂犬病病毒(PRV)、猪繁殖与呼吸综合征病毒(PRRSV)、猪瘟病毒(CSFV)等猪常见病原的检测结果均为阴性;与普通PCR方法相比,其敏感性高出100倍,可达100拷贝/μL;对临床样品的检测证明,该方法可有效检测出淋巴结、肺脏等组织中的PCV2。The research with porcine circovirus type 2 （PCV2） of the ORF2 capsid protein gene for the purpose, design and synthesis of specific primers and probes. Gene was amplified by PCR and cloned into the pMD18-T vector, screened positive plasmid standards. By quantitative PCR optimization of reaction conditions, establishment of a real-time detection of PCV2 Taq Man quantitative PCR method. The results showed that the method was specificity, the pseudorabies virus （PRV）, porcine reproductive and respiratory syndrome virus （PRRSV）, swine fever virus （CSFV） and other common diseases of the original pig detection results were negative. Compared with ordinary PCR method, the sensitivity was 100 times higher, up to 100 cop- ies/μL; for the detection of clinical samples showed that the method can effectively detect lymph nodes, lungs and other tissues of PCV2.

关 键 词：猪圆环病毒2型 荧光定量PCR TAQMAN探针 标准曲线 

分 类 号：S852.659.2[农业科学—基础兽医学]