Peng Yadan, Hu Ruolan, Xue Shuang, He Yugan, Tian Lili, Pang Zehan, He Yile, Dong Yuqi, Shi Yinghan, Wang Shuqi, Hong Bixia, Liu Ke, Wang Ruixue, Song Lihua, Fan Huahao, Li Mengzhe, Tong Yigang
College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, 100029, China.
College of Mechanical and Electrical Engineering, Beijing University of Chemical Technology, Beijing, 100029, China.
Anal Chim Acta. 2024 Jul 4;1311:342720. doi: 10.1016/j.aca.2024.342720. Epub 2024 May 14.
The monkeypox virus (MPXV) is a linear double-stranded DNA virus with a large genome that causes tens of thousands of infections and hundreds of deaths in at least 40 countries and regions worldwide. Therefore, timely and accurate diagnostic testing could be an important measure to prevent the ongoing spread of MPXV and widespread epidemics.
Here, we designed multiple sets of primers for the target region of MPXV for loop-mediated isothermal amplification (LAMP) detection and identified the optimal primer set. Then, the specificity in fluorescent LAMP detection was verified using the plasmids containing the target gene, pseudovirus and other DNA/RNA viruses. We also evaluated the sensitivity of the colorimetric LAMP detection system using the plasmid and pseudovirus samples, respectively. Besides, we used monkeypox pseudovirus to simulate real samples for detection. Subsequent to the establishment and introduction of a magnetic beads (MBs)-based nucleic acid extraction technique, an integrated device was developed, characterized by rapidity, high sensitivity, and remarkable specificity. This portable system demonstrated a visual detection limit of 137 copies/mL, achieving sample-to-answer detection within 1 h.
The device has the advantages of integration, simplicity, miniaturization, and visualization, which help promote the realization of accurate, rapid, portable, and low-cost testing. Meanwhile, this platform could facilitate efficient, cost-effective and easy-operable point-of-care testing (POCT) in diverse resource-limited settings in addition to the laboratory.
猴痘病毒（MPXV）是一种线性双链DNA病毒，基因组庞大，在全球至少40个国家和地区导致了数以万计的感染和数百人死亡。因此，及时准确的诊断检测可能是预防猴痘病毒持续传播和广泛流行的一项重要措施。
在此，我们针对猴痘病毒的靶区域设计了多组引物用于环介导等温扩增（LAMP）检测，并确定了最佳引物组。然后，使用含有靶基因的质粒、假病毒及其他DNA/RNA病毒验证了荧光LAMP检测的特异性。我们还分别使用质粒和假病毒样本评估了比色LAMP检测系统的灵敏度。此外，我们使用猴痘假病毒模拟真实样本进行检测。在建立并引入基于磁珠（MBs）的核酸提取技术后，开发了一种集成装置，其特点是快速、高灵敏度和显著的特异性。这个便携式系统的目视检测限为137拷贝/毫升，在1小时内实现了样本到结果的检测。
该装置具有集成化、简单、小型化和可视化的优点，有助于推动实现准确、快速、便携和低成本检测。同时，除实验室外，该平台还可在各种资源有限的环境中促进高效、经济高效且易于操作的即时检测（POCT）。
