Rapid detection of botulinum toxins using a high sensitivity SPR sensor

使用高灵敏度 SPR 传感器快速检测肉毒杆菌毒素

• 批准号: 7924013
• 负责人: PAUL MELMAN
• 金额: $ 50.23万
• 依托单位: NEWTON PHOTONICS
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7924013
• 关键词: Affinity; Animal Model; Antitoxins; Biological; Biological Assay; Blood; Bontoxilysin; Botulinum Antitoxin; Botulinum Toxin Type A; Botulinum Toxins; Botulism; Clinical; Data; Detection; Diagnosis; Diagnostic; Early Diagnosis; Exposure to; Goals; Gold; Half-Life; Health care facility; Human; Human Resources; In Vitro; Individual; Intoxication; Laboratories; Laboratory Animals; Laboratory Diagnosis; Metabolism; Methods; Mus; Nerve Endings; Peptides; Poisoning; Protocols documentation; Public Health; Sampling; Serotyping; Serum; Surface Plasmon Resonance; Symptoms; Technology; Time; Toxin; Training; base; biothreat; botulinum; botulinum toxin type B; nanoparticle; peptide A; rapid detection; sensor; weapons

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.

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。

项目成果

Comparative immunochemical characteristics of botulinum neurotoxin type A and its associated proteins.

• DOI: 10.1016/j.toxicon.2013.06.011
• 发表时间: 2013-09
• 期刊: Toxicon : official journal of the International Society on Toxinology
• 作者: Bryant AM;Cai S;Singh BR
• 通讯作者: Singh BR

High Yield Preparation of Functionally Active Catalytic-Translocation Domain Module of Botulinum Neurotoxin Type A That Exhibits Uniquely Different Enzyme Kinetics.

高产地制备具有独特不同酶动力学的 A 型肉毒杆菌神经毒素功能活性催化转位结构域模块。

• DOI: 10.1007/s10930-017-9744-8
• 发表时间: 2017
• 期刊: The protein journal
• 作者: Dhaliwal,HarkiranpreetKaur;Thiruvanakarasu,Nagarajan;Kumar,Raj;Patel,Kruti;Ambrin,Ghuncha;Cai,Shouwei;Singh,BalRam
• 通讯作者: Singh,BalRam