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PRRSV和PCV-2以及PRV多重SYBR
Green-Ⅰ实时荧光PCR检测方法的建立
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王小武1，符芳1，柴政1，孔令达1，蔡雪辉1，宋淑萍1，许红喜2，李曦1*
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（1.中国农业科学院 哈尔滨兽医研究所 兽医生物技术国家重点实验室 猪传染病研究室,黑龙江
哈尔滨150001；2.黑龙江八一农垦大学,黑龙江 大庆163319）
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摘要：
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根据GenBank中登录的猪生殖与呼吸综合征病毒(PRRSV) N蛋白基因、猪圆环病毒2型(PCV-2) rep蛋白基因和猪伪狂犬病病毒(PRV) gE基因的核苷酸序列分别设计了3对特异性引物，成功建立了同时检测PRRSV、PCV-2、PRV的多重SYBR Green-Ⅰ实时荧光PCR方法。敏感性试验结果显示，PRRSV、PCV-2的敏感性可达250拷贝/μL，PRV的敏感性可达500拷贝/μL。表明，该方法具有较好的特异性、重复性和敏感性，可以用于PRRSV、PCV-2和PRV的快速检测。 |
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关键词：
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猪生殖与呼吸综合征病毒；猪圆环病毒2型；猪伪狂犬病病毒；多重SYBR Green-Ⅰ实时荧光PCR |
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中图分类号：S 852.659.6
文献标识码：
A 文章编号：1673-4696(2008)06-0494-06 |
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Establishment of a multiplex SYBR Green-Ⅰ real time PCR assay for
simultaneous detection of porcine reproductive and respiratory
syndrome virus,porcine circovirus type 2
and porcine pseudorabies virus |
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WANG Xiao-wu1,FU Fang1,CHAI Zheng1,KONG
Ling-da1,CAI Xue-hui1,SONG Shu-ping1,
XU Hong-xi2,LI Xi1 |
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(1.National Key Laboratory of Veterinary Biotechnology/Division of Swine Infectious Diseases,Harbin
Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150001,
China;2.Heilongjiang August-First Land Reclamation University,Daqing 163319,China) |
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Abstract: Three pairs of primers were designed respectively based on the sequences of N protein gene of PRRSV,rep protein gene of PCV-2 and gE gene of PRV available in GenBank.A multiplex SYBR Green-Ⅰ real time PCR assay was established successfully for simultaneous detection of the 3 viruses.The sensitivities for PRRSV,PCV-2 and PRV were 250,250 and 500 copies/μL,respectively.The results revealed that the established PCR assay was sensitive,specific and reproducible and it could be used to detect rapidly PRRSV,PCV-2 and PRV. |
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Key words: porcine reproductive and respiratory syndrome virus;porcine circovirus type 2;porcine pseudorabies virus;multiplex SYBR Green-Ⅰ real time PCR |
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