[1]季程远,王蔚怡,黄 欣,等. 猪圆环病毒2型和3型双重PCR检测方法的建立与应用[J].中国预防兽医学报,2018,(10):913-916.[doi:10.3969/j.issn.1008-0589.201801022]
 JI Cheng-yuan,WANG Wei-yi,HUANG Xin,et al. Establishment and application of a duplex PCR for detection of porcine circovirus 2 and porcine circovirus 3[J].Chinese journal of preventive veterinary medicine,2018,(10):913-916.[doi:10.3969/j.issn.1008-0589.201801022]
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猪圆环病毒2型和3型双重PCR检测方法的建立与应用

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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2018年10
页码:
913-916
栏目:
诊断和检测技术
出版日期:
2018-11-15

文章信息/Info

Title:
 

Establishment and application of a duplex PCR for detection of porcine circovirus 2 and porcine circovirus 3

文章编号:
1008-0589(2018)10-0913-04
作者:
 

季程远王蔚怡黄 欣方庆励欧阳康陈 樱韦祖樟黄伟坚*

 

(广西大学 动物科学技术学院,广西 南宁 530005)

Author(s):
 

JI Cheng-yuan WANG Wei-yi HUANG Xin FANG Qing-li OUYANG Kang

CHEN Ying WEI Zu-zhang HUANG Wei-jian*

 

(College of Animal Science and Technology, Guangxi University, Nanning 530005, China)

关键词:
猪圆环病毒2型猪圆环病毒3型双重PCR检测
Keywords:
 

 PCV2 PCV3 duplex PCR detection

分类号:
S852.65
DOI:
10.3969/j.issn.1008-0589.201801022
文献标志码:
A
摘要:
 

为建立一种快速且同时检测猪圆环病毒2型(PCV2)和3型(PCV3)的方法,本研究根据GenBank中PCV2和 PCV3的全基因组序列,设计了两对特异性引物,通过对扩增条件的优化,建立了能够同时检测PCV2和PCV3的双重PCR方法。该方法可以同时扩增PCV2的266 bp和PCV3 的594 bp特异性片段,而对猪伪狂犬病毒、猪细小病毒、猪瘟病毒、猪繁殖障碍与呼吸综合征病毒和猪流行性腹泻病毒的基因组扩增结果均为阴性。对PCV2和PCV3的最低检出量均为100拷贝/μL。对广西境内100份健康屠宰猪淋巴结样品检测结果显示,单一PCR与双重PCR检测符合率为100 %。结果表明,本研究建立了一种简便快捷、反应灵敏、稳定可靠且特异性强的双重PCR技术,为临床样品中PCV2和PCV3的快速检测提供了有效的技术支持。

  

Abstract:
 

To develop a rapid duplex PCR assay for simultaneous detection of porcine circuvirus 2 (PCV2) and porcine circuvirus 3(PCV3), two pairs of specific primers was designed based on the published sequence of the complete genome of PCV2 and PCV3 in GenBank. The duplex PCR was successfully established by optimizing the duplex PCR amplification conditions. The specific PCR products of 266bp for PCV2 and 594bp for PCV3 were simultaneously amplified from the DNAs of PCV2 and PCV3 by the duplex PCR, but no PCR products were amplified from porcine parvovirus, pseudorabies virus, swine fever virus, porcine reproductive and respiratory syndrome virus, and porcine epidemic diarrhea virus. The duplex PCR method was capable of detecting the template DNA of 100copies/μL for both PCV2 and PCV3. The coincidence between single PCR assays and duplex PCR was 100% by detecting 100 lymph node samples. The results suggest that the duplex PCR was characterized by convenience, high sensitivity, stability and specificity, which provides effective support for differentiating the clinical samples with PCV2 and PCV3 co-infection.

参考文献/References:

 

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(本文编辑:李 娜)

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更新日期/Last Update: 2018-11-28