Detection of botulinum neurotoxin-A activity in food by peptide cleavage assay
Detection of botulinum neurotoxin-A activity in food by peptide cleavage assay
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DOI：
10.1016/j.ijfoodmicro.2008.05.012
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发表时间：
2008-08-15
影响因子：
5.4
通讯作者：
Brandon, David L.
中科院分区：
文献类型：
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作者：
Rasooly, Reuven;Stanker, Larry H.;Brandon, David L.
世界卫生组织 (WHO) 和美国疾病控制与预防中心 (CDC) 已将肉毒杆菌毒素标记为高度优先的生物制剂，可用于针对食品供应的恐怖袭击。由于这种威胁，越来越需要开发快速有效的方法来检测活性肉毒杆菌神经毒素（BoNT）。这项研究报告了使用内部淬灭荧光肽的酶测定法的成功应用，作为小鼠生物测定法的快速、简单且廉价的替代方案。不到 15 分钟，该检测即可检测液体食品样品中的 0.25 nM BoNT-A。检出水平远低于成人口服致死剂量70微克毒素。包被有针对毒素重链的IgG单克隆抗体的免疫磁珠可以浓缩毒素而不中和其酶活性，克服内源性蛋白酶抑制剂和肽酶引起的基质效应。这种快速有效的检测系统可用于大规模筛查以检测 BoNT-A。由 Elsevier B.V. 出版
The World Health Organization (WHO) and U.S. Centers for Disease Control and Prevention (CDC) have labeled botulinum toxins as a high priority biological agent that may be used in terrorist attacks against food supplies. Due to this threat there is an increased need to develop fast and effective methods to detect active botulinum neurotoxins (BoNTs). This study reports the successful use of an enzymatic assay employing an internally quenched fluorogenic peptide as a fast, simple and inexpensive alternative to the mouse bioassay. In less than 15 min the assay can detect 0.25 nM BoNT-A in liquid food samples. The detection level is far below the adult human lethal oral dose of 70 mu g of toxin. Immunomagnetic beads coated with IgG monoclonal antibodies that target the toxin heavy chain can concentrate the toxin without neutralizing its enzymatic activity, overcoming matrix effects caused by endogenous protease inhibitors and peptidases. This fast and effective assay system could be used for large scale screening to detect BoNT-A. Published by Elsevier B.V.