检测猪圆环病毒2型SYBRGreenⅠ荧光定量PCR方法的建立  

Establishment of SYBR Green Ⅰ Real-time PCR Aaasy for Detection of Porcine Circovirus Type 2

作　　者：张锦秀[1] 晁生玉[1] 丁敏[1] 李刚[1] 

机构地区：[1]中国农业科学院北京畜牧兽医研究所,北京100193

出　　处：《中国畜牧兽医》2008年第9期30-33,共4页China Animal Husbandry & Veterinary Medicine

基　　金：中央级公益性科研院所基本科研业务费专项(0032007008)

摘　　要：猪圆环病毒2型(PCV-2)ORF2(开放阅读框2)基因编码其核衣壳蛋白,它含有导致病毒致病性的多个抗原决定簇位点。根据ORF2基因序列设计特异性引物,以感染PCV-2的Dulac细胞冻融上清为模板,克隆该开放阅读框的全基因序列。然后以该重组质粒为模板,选择其异于PCV-1的保守序列,设计PCR引物,采用两步法,建立了检测PCV-2快速简捷的SYBR Green Ⅰ荧光定量PCR方法。多次试验证明,该方法具有良好的重复性。同时针对与PCV-2共感染的其它病毒进行特异性检测,进一步证明该方法可以快速、特异、灵敏地检测PCV-2。Porcine circovirus type 20RF2 encodes the capsid protein which contains major immunoreactive epitopes. The whole sequence of the ORF2 was amplified with a special pair of primers and cloned into vectors. The PCV-2 DNA was obtained from the supernatant of PCV-2 infected Dulac cells after freeze thrawing. Then a two-step method of SYBR Green I Real-time PCR amplification was established for the rapid and sensitive detection of PCV-2. The detection results confirmed that the es- tablished method was rapid, specific and sensitive.

关 键 词：猪圆环病毒2型 Dulac细胞 SYBR Green Ⅰ荧光定量PCR 

分 类 号：S852.659.2[农业科学—基础兽医学]