Shandong Provincial Key Laboratory of Animal Disease Control & Breeding, Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan, China.
Qingdao Agricultural University, Qingdao, China.
Transbound Emerg Dis. 2018 Jun;65(3):597-601. doi: 10.1111/tbed.12835. Epub 2018 Mar 4.
In this study, a rapid and specific assay for the detection of porcine circovirus type 3 (PCV3) was established using loop-mediated isothermal amplification (LAMP). Four primers were specifically designed to amplify PCV3. The LAMP assay was effectively optimized to amplify PCV3 by water bath at 60°C for 60 min. The detection limit was approximately 1 × 10 copy in this LAMP assay. Compared to porcine circovirus type 2 (PCV2), both gE and gD genes of pseudorabies virus (PRV) and porcine parvovirus (PPV), the LAMP assay showed a high specific detection of PCV3. A visible detection method was developed using SYBR Green I to recognize the results rapidly. Based on the detection of 20 clinical tissue samples, the LAMP assay was more practical and convenient than classical PCR due to its simplicity, high sensitivity, rapidity, specificity, visibility and cost efficiency.
在这项研究中，我们建立了一种使用环介导等温扩增（LAMP）技术快速且特异性检测猪圆环病毒 3 型（PCV3）的方法。我们专门设计了四个引物来扩增 PCV3。通过优化 LAMP 反应，在 60°C 的水浴中孵育 60 分钟即可有效地扩增 PCV3。该 LAMP 检测方法的检测限约为 1×10 拷贝。与猪圆环病毒 2 型（PCV2）相比，该方法对伪狂犬病病毒（PRV）和猪细小病毒（PPV）的 gE 和 gD 基因均具有较高的特异性检测。我们使用 SYBR Green I 开发了一种可见的检测方法，可快速识别结果。基于对 20 个临床组织样本的检测，LAMP 检测方法由于其简单、高灵敏度、快速、特异性、可视化和成本效益，比经典 PCR 更实用和方便。
