Rapid detection of botulinum toxins using a high sensitivity SPR sensor

描述（由申请人提供）：肉毒杆菌中毒是一种严重且公认的生物威胁。肉毒杆菌神经毒素（BoNT）非常有效，相对容易制备/支付，并且已经被用作生物恐怖剂。在临床症状发展到临界状态之前，迫切需要一种敏感和特异性的方法来早期检测暴露个体血液中的BoNT。目前肉毒杆菌中毒实验室诊断的“金标准”是小鼠生物测定法。虽然敏感，但速度也很慢（2-4天），需要专门的动物实验室。这凸显了对快速、敏感和易于使用的BoNT检测技术的需求。该项目的总体目标是建立一种快速（30~60分钟）体外检测血清中BoNT/ a和BoNT/B的传感器。该传感器将用于诊断实验室和公共卫生设施。该传感器将基于表面等离子体共振技术的成熟波导实现而构建。它将使用酶切法测定BoNT/A和BoNT/B。合成一个肽底物，包括BoNT/A和BoNT/B的裂解序列。金纳米颗粒会附着在肽上，以提高SPR检测的灵敏度。将开发一种基于免疫亲和力的样品方案，从血清中提取和浓缩目标bont。

DESCRIPTION (provided by applicant): Botulinum poisoning is a serious and well-recognized biothreat. Botulinum neurotoxins (BoNT) are extremely potent, relatively simple to prepare/disburse, and have already been used as a bioterror agent. There is a vital need for sensitive and specific methods for early detection of BoNT in the blood of exposed individuals before clinical symptoms have progressed to a critical state. The current "gold standard" for laboratory diagnosis of botulism is the mouse bioassay. While sensitive, it is also slow (2-4 days) and requires a dedicated animal laboratory. This underscores the unmet need for a rapid, sensitive and easy to use BoNT detection technology. The overall goal of the project is to build a sensor for rapid (30~60 minutes) in-vitro detection of BoNT/A and BoNT/B in serum. The sensor will be aimed at use in diagnostic laboratories and public health facilities. The sensor will be built based on a proven waveguide implementation of the surface plasmon resonance technology. It will use enzymatic cleavage assays for BoNT/A and BoNT/B. A peptide substrate will be synthesized, including the cleavage sequences for BoNT/A and BoNT/B. Gold nanoparticles will be attached to the peptide to increase the SPR detection sensitivity. An immuno-affinity based sample protocol will be developed to extract and concentrate the target BoNTs from serum.