Rapid detection of porcine circovirus type 2 by loop-mediated isothermal amplification
- 作者：Hao-tai Chen ; Jie Zhang ; De-hui Sun ; Yue-feng Chu ; Xue-peng Cai ; Xiang-tao Liu ; Xue-nong Luo ; Qing Liu ; Yong-sheng Liu
- 关键词：Porcine circovirus type 2 (PCV2) ; Detection ; Loop-mediated isothermal amplification (LAMP) ; Sensitivity ; Specificity
- 刊名：Journal of Virological Methods
- 出版年：2008
- 期刊代码：180_01660934
- 类别：med
- 出版时间：May 2008
- 卷：149
- 期：2
- 页码：264-268
- 文件大小：197 K
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 °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.