Jenko Kathryn L, Zhang Yanfeng, Kostenko Yulia, Fan Yongfeng, Garcia-Rodriguez Consuelo, Lou Jianlong, Marks James D, Varnum Susan M
Biological Sciences Division, Pacific Northwest National Laboratory, Richland, WA 99354, USA.
Analyst. 2014 Oct 21;139(20):5093-102. doi: 10.1039/c4an01270d. Epub 2014 Aug 12.
Plant and microbial toxins are considered bioterrorism threat agents because of their extreme toxicity and/or ease of availability. Additionally, some of these toxins are increasingly responsible for accidental food poisonings. The current study utilized an ELISA-based protein antibody microarray for the multiplexed detection of ten biothreat toxins, botulinum neurotoxins (BoNT) A, B, C, D, E, F, ricin, shiga toxins 1 and 2 (Stx), and staphylococcus enterotoxin B (SEB), in buffer and complex biological matrices. The multiplexed assay displayed a sensitivity of 1.3 pg mL(-1) (BoNT/A, BoNT/B, SEB, Stx-1 and Stx-2), 3.3 pg mL(-1) (BoNT/C, BoNT/E, BoNT/F) and 8.2 pg mL(-1) (BoNT/D, ricin). All assays demonstrated high accuracy (75-120 percent recovery) and reproducibility (most coefficients of variation <20%). Quantification curves for the ten toxins were also evaluated in clinical samples (serum, plasma, nasal fluid, saliva, stool, and urine) and environmental samples (apple juice, milk and baby food) with overall minimal matrix effects. The multiplex assays were highly specific, with little cross-reactivity observed between the selected toxin antibodies. The results demonstrate a multiplex microarray that improves current immunoassay sensitivity for biological warfare agents in buffer, clinical, and environmental samples.
植物和微生物毒素因其极高的毒性和/或易于获取而被视为生物恐怖主义威胁因子。此外，其中一些毒素导致意外食物中毒的情况日益增多。当前的研究利用基于酶联免疫吸附测定（ELISA）的蛋白质抗体微阵列，对缓冲液和复杂生物基质中的十种生物威胁毒素进行多重检测，这些毒素包括肉毒杆菌神经毒素（BoNT）A、B、C、D、E、F、蓖麻毒素、志贺毒素1和2（Stx）以及葡萄球菌肠毒素B（SEB）。多重检测显示出的灵敏度为：1.3 pg mL⁻¹（BoNT/A、BoNT/B、SEB、Stx - 1和Stx - 2）、3.3 pg mL⁻¹（BoNT/C、BoNT/E、BoNT/F）以及8.2 pg mL⁻¹（BoNT/D、蓖麻毒素）。所有检测均显示出高准确度（回收率75 - 120%）和可重复性（大多数变异系数<20%）。还在临床样本（血清、血浆、鼻液、唾液、粪便和尿液）和环境样本（苹果汁、牛奶和婴儿食品）中评估了这十种毒素的定量曲线，总体基质效应最小。多重检测具有高度特异性，在所选用的毒素抗体之间几乎未观察到交叉反应。结果表明，这种多重微阵列提高了当前免疫测定对缓冲液、临床和环境样本中生物战剂的灵敏度。
