Multiplex PCR for rapid detection of pseudorabies virus,porcine parvovirus and porcine circovirus type 2

PAN Qun-xing; CHEN De; HE Kong-wang; HUANG Ke-he

PDF
Cite
Share

facebook

twitter

google

linkedin

December 2005

PAN Qun-xing, CHEN De, HE Kong-wang and HUANG Ke-he. Multiplex PCR for rapid detection of pseudorabies virus,porcine parvovirus and porcine circovirus type 2[J]. Virologica Sinica, 2005, 20(6): 603-606.

Citation: PAN Qun-xing, CHEN De, HE Kong-wang, HUANG Ke-he. Multiplex PCR for rapid detection of pseudorabies virus,porcine parvovirus and porcine circovirus type 2 .VIROLOGICA SINICA, 2005, 20(6) : 603-606.

检测PCV_2、PPV、PRV疫苗株与野毒株的多重PCR方法

潘群兴 ^1,2 ,
陈德 ^1,2 ,
何孔旺 ^2* ,
黄克和 ^1,,

1.
1. 江苏南京江苏省农业科学院兽医研究所 2.农业部畜禽疫病诊断重点开放实验室南京农业大学动物医学院

通讯作者： 黄克和,

摘要

本文建立了一种同时检测猪圆环病毒2型(PCV2)、细小病毒(PPV)、及伪狂犬病毒(PRV)疫苗株与野毒株的多重PCR方法。根据GenBank上发表的PCV2、PPV和PRV gB、gE基因序列,针对各自保守区各设计一对特异性引物,用这四对引物对同一样品中的PCV2、PPV和PRV gB、gE进行检测,结果可同时扩增出269bp(PCV2)、 581bp(PPV)、372bP(PRV gB)及147bp(PRV gE)四条特异性片段。对JEV、PRRSRV、大肠杆菌和双蒸水的PCR 扩增结果均为阴性;敏感性测定结果表明,该多重PCR能检出10pg PCV2、PPV和PRV gB、gE检测敏感度分别为 106.2、103.8、10-5.8 TCID50的模板。该方法的建立对临床上进行这三种疾病的鉴别诊断和混合感染的检测具有重要意义。
- 多重聚合酶链式反应
- , 伪狂犬病毒（PRV）
- , 细小病毒（PPV）
- , 猪圆环病毒2型（PCV2）

Multiplex PCR for rapid detection of pseudorabies virus,porcine parvovirus and porcine circovirus type 2

PAN Qun-xing ^1,2 ,
CHEN De ^1,2 ,
HE Kong-wang ^2* ,
HUANG Ke-he ^1,,

Corresponding author: HUANG Ke-he,

Abstract

A multiplex PCR(mPCR) assay was developed and evaluated for its effectiveness as a means to simultaneously detect multiple viral infection of swine . Specific primers for each of the three common DNA viruses, Pseudorabies virus(PRV) , Porcine parvovirus ( PPV) and Porcine circovirus type 2(PCV2) were used to test the procedure, Four specific bands of 269bp(PCV2), 583 bp(PPV) 372 bp(PRV gB)and147 bp(PRV gE)were amplified. The assay proved to be sensitive when a composite of all three viruses were amplified, including both field and gene-deleted permutations of PRV. No specific band was amplified from other pathogenic viruses and bacteria. As little as 10 pg PCV2, 10-6.2 TCID50 PPV,10-3.8 TCID50\PRV gB and 10-5.8 TCID50 PRV gE were detected in this mPCR. This method could effectively detect infection of PCV2, PPV, field and gene-deleted permutations of PRV from clinical samples.
- Multiplex
- , PCR Pseudorabies virus（PRV）
- , Porcine parvovirus （PPV）
- , Porcine circo-virus type 2（PCV2）

References
Proportional views

PDF

Article Metrics

Article views(4799) PDF downloads(781) Cited by(0)

Proportional views

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Multiplex PCR for rapid detection of pseudorabies virus,porcine parvovirus and porcine circovirus type 2

Corresponding author: HUANG Ke-he,

Abstract: A multiplex PCR(mPCR) assay was developed and evaluated for its effectiveness as a means to simultaneously detect multiple viral infection of swine . Specific primers for each of the three common DNA viruses, Pseudorabies virus(PRV) , Porcine parvovirus ( PPV) and Porcine circovirus type 2(PCV2) were used to test the procedure, Four specific bands of 269bp(PCV2), 583 bp(PPV) 372 bp(PRV gB)and147 bp(PRV gE)were amplified. The assay proved to be sensitive when a composite of all three viruses were amplified, including both field and gene-deleted permutations of PRV. No specific band was amplified from other pathogenic viruses and bacteria. As little as 10 pg PCV2, 10-6.2 TCID50 PPV,10-3.8 TCID50\PRV gB and 10-5.8 TCID50 PRV gE were detected in this mPCR. This method could effectively detect infection of PCV2, PPV, field and gene-deleted permutations of PRV from clinical samples.