Rapid detection of porcine circovirus type 2 by loop-mediated isothermal amplification

被引：52

作者：

Chen, Hao-tai ^{[1
]}

Zhang, Jie ^{[1
]}

Sun, De-hui ^{[1
]}

Chu, Yue-feng ^{[1
]}

Cai, Xue-peng ^{[1
]}

Liu, Xiang-tao ^{[1
]}

Luo, Xue-nong ^{[1
]}

Liu, Qing ^{[2
]}

Liu, Yong-sheng ^{[1
]}

机构：

[1] Chinese Acad Agr Sci, Lanzhou Vet Res Inst, Dept Virol,Minist Agr, State Key Lab Vet Etiol Biol,Key Lab Anim Virol, Lanzhou 730046, Gansu, Peoples R China

[2] Chinese Acad Sci, Inst Modern Phys, Lanzhou 730046, Peoples R China

来源：

JOURNAL OF VIROLOGICAL METHODS | 2008年 / 149卷 / 02期

基金：

中国国家自然科学基金;

关键词：

porcine circovirus type 2 (PCV2); detection; loop-mediated isothermal amplification (LAMP); sensitivity; specificity;

D O I：

10.1016/j.jviromet.2008.01.023

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A method of loop-mediated isothermal amplification (LAMP) was employed to develop a rapid and simple detection system for porcine circovirus type 2 (PCV2). The amplification could be finished in 60 min under isothermal condition at 64 degrees C by employing a set of four primers targeting the cap gene of PCV2. The LAMP assay showed higher sensitivity than the conventional PCR, with a detection limit of five copies per tube of purified PCV2 genomic DNA. No cross-reactivity was observed from the samples of other related viruses including porcine circovirus type 1 (PCV1), porcine parvovirus (PPV), porcine pseudorabies virus (PRV) and porcine reproductive and respiratory syndrome virus (PRRSV). The detection rate of PCV2 LAMP for 86 clinical samples was 96.5% and appeared greater than that of the PCR method. The LAMP assay reported can provide a rapid yet simple test of PCV2 suitable for laboratory diagnosis and pen-side detection due to ease of operation and the requirement of only a regular water bath or heat block for the reaction. (c) 2008 Elsevier B.V. All rights reserved.

引用

页码：264 / 268

页数：5

共 33 条

[1] Isolation of porcine circovirus-like viruses from pigs with a wasting disease in the USA and Europe [J].