Liu Jing, Gao Shan, Kang Lin, Ji Bin, Xin Wenwen, Kang Jingjing, Li Ping, Gao Jie, Wang Hanbin, Wang Jinglin, Yang Hao
State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Beijing, 100071, People's Republic of China.
The 307th Hospital of Military Chinese People's Liberation Army, Beijing, 100071, People's Republic of China.
Nanoscale Res Lett. 2017 Dec;12(1):227. doi: 10.1186/s11671-017-1944-9. Epub 2017 Mar 29.
Botulism is a severe and potentially lethal paralytic disease caused by several botulinum neurotoxin-producing Clostridia spp. In China, the majority of the cases caused by botulism were from less-developed rural areas. Here, we designed specific substrate peptides and reconfigured gold nanoparticle-based lateral flow test strip (LFTS) to develop an endopeptidase-based lateral flow assay for the diagnosis of botulism. We performed this lateral flow assay on botulinum neurotoxin-spiked human serum samples. The as-prepared LFTS had excellent performance in the detection of botulinum neurotoxin using only 1 μL of simulated serum, and its sensitivity and specificity were comparable to that of mouse lethality assay. Moreover, the assay takes only half a day and does not require highly trained laboratory staff, specialized facility, or equipment. Finally, our LFTS can be potentially extended to other serotypes of BoNTs by designing specific substrate peptides against the different types of BoNTs. Overall, we demonstrate a strategy by which LFTS and endopeptidase activity assays can be integrated to achieve facile and economic diagnosis of botulism in resource-limited settings.
肉毒中毒是由几种产生肉毒杆菌神经毒素的梭菌属细菌引起的一种严重且可能致命的麻痹性疾病。在中国，大多数肉毒中毒病例来自欠发达的农村地区。在此，我们设计了特定的底物肽并重新配置了基于金纳米颗粒的侧向流动试纸条（LFTS），以开发一种基于内肽酶的侧向流动分析法用于肉毒中毒的诊断。我们在添加了肉毒杆菌神经毒素的人血清样本上进行了这种侧向流动分析。所制备的LFTS仅使用1μL模拟血清就能在检测肉毒杆菌神经毒素方面表现出优异的性能，其灵敏度和特异性与小鼠致死率测定法相当。此外，该分析仅需半天时间，且不需要训练有素的实验室工作人员、专门的设施或设备。最后，通过针对不同类型的肉毒杆菌神经毒素设计特定的底物肽，我们的LFTS有可能扩展到其他血清型的肉毒杆菌神经毒素。总体而言，我们展示了一种策略，通过该策略可以将LFTS和内肽酶活性分析相结合，以在资源有限的环境中实现肉毒中毒的简便且经济的诊断。
