Zhu Yuan-Shou, Shao Ning, Chen Jian-Wei, Qi Wen-Bao, Li Yang, Liu Peng, Chen Yan-Jing, Bian Su-Ying, Zhang Yan, Tao Sheng-Ce
Shanghai Center for Systems Biomedicine, Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Jiao Tong University, Shanghai 200240, China.
College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China.
Anal Chim Acta. 2020 Dec 15;1140:30-40. doi: 10.1016/j.aca.2020.10.011. Epub 2020 Oct 8.
African swine fever is caused by African swine fever virus (ASFV), and has a mortality rate approaching 100%. It has already caused tremendous economy lost around the world. Without effective vaccine, rapid and accurate on-site detection plays an indispensable role in controlling outbreaks. Herein, by combining Hive-Chip and direct loop-mediated isothermal amplification (LAMP), we establish a multiplex and visual detection platform. LAMP primers targeting five ASFV genes (B646L, B962L, C717R, D1133L, and G1340L) were designed and pre-fixed in Hive-Chip. On-chip LAMP showed the limits of detection (LOD) of ASFV synthetic DNAs and mock samples are 30 and 50 copies per microliter, respectively, and there is no cross-reaction among the target genes. The overall performance of our platform is comparable to that of the commercial kits. From sample preparation to results readout, the entire process takes less than 70 min. Multiplex detection of real samples of ASFV and other swine viruses further demonstrates the high sensitivity and specificity of Hive-Chip. Overall, our platform provides a promising option for on-site, fast and accurate detection of ASFV.
非洲猪瘟由非洲猪瘟病毒（ASFV）引起，死亡率接近100%。它已在全球范围内造成了巨大的经济损失。由于没有有效的疫苗，快速准确的现场检测在控制疫情爆发中起着不可或缺的作用。在此，通过将蜂巢芯片（Hive-Chip）与直接环介导等温扩增（LAMP）相结合，我们建立了一个多重可视化检测平台。针对五个ASFV基因（B646L、B962L、C717R、D1133L和G1340L）设计的LAMP引物被预固定在蜂巢芯片中。芯片上的LAMP显示，ASFV合成DNA和模拟样本的检测限（LOD）分别为每微升30拷贝和50拷贝，并且目标基因之间没有交叉反应。我们平台的整体性能与商业试剂盒相当。从样品制备到结果读取，整个过程耗时不到70分钟。对ASFV和其他猪病毒的实际样本进行多重检测进一步证明了蜂巢芯片的高灵敏度和特异性。总体而言，我们的平台为ASFV的现场快速准确检测提供了一个有前景的选择。
