Animal Health Laboratory, Indian Council of Agricultural Research, National Research Centre on Pig (ICAR- NRC on Pig), Assam, India.
Department of Microbiology, AAU, Khanapara, India.
Anim Biotechnol. 2023 Dec;34(7):2441-2448. doi: 10.1080/10495398.2022.2095516. Epub 2022 Jul 6.
A cost effective, simple and rapid method is critical for detection of porcine circovirus type 2 (PCV2) infection in pigs. The present study reports the development and evaluation a loop-mediated isothermal amplification (LAMP) assay for rapid visual detection of PCV2 in pigs. The time and temperature conditions for amplification of PCV2 genes were optimized to be 30 min at 67 °C. The developed assay was 10 fold more sensitive than conventional PCR with analytical sensitivity of 5 pg and 50 pg, respectively. The developed LAMP assay had a sensitivity of 100%, specificity of 85.45% and overall accuracy of 89.70%. This is perhaps the most rapid of all LAMP reports for PCV2 detection available globally. The assay did not cross-react with porcine parvovirus or classical swine fever virus. DNA sequencing was done to ensure accuracy of LAMP assay results. The assay was assembled into a kit of 20 reactions and validated in different laboratories in India. The developed LAMP assay was proved to be a specific, sensitive and rapid method for visual detection of PCV2 which does not require costly equipments.
一种经济有效、简单快速的方法对于检测猪圆环病毒 2 型（PCV2）感染至关重要。本研究报告了一种用于快速目视检测猪 PCV2 的环介导等温扩增（LAMP）检测方法的开发和评估。优化了用于扩增 PCV2 基因的时间和温度条件，使其在 67°C 下 30 分钟即可完成扩增。与传统 PCR 相比，该方法的灵敏度提高了 10 倍，分别为 5pg 和 50pg。该开发的 LAMP 检测方法的灵敏度为 100%，特异性为 85.45%，总准确率为 89.70%。这也许是全球所有 PCV2 检测中最快的 LAMP 报告之一。该检测方法与猪细小病毒或古典猪瘟病毒没有交叉反应。进行 DNA 测序以确保 LAMP 检测结果的准确性。该检测方法被组装成 20 个反应试剂盒，并在印度的不同实验室进行了验证。开发的 LAMP 检测方法被证明是一种用于目视检测 PCV2 的特异性、灵敏和快速方法，它不需要昂贵的设备。
