Alam S I, Uppal A, Gupta P, Kamboj D V
Biotechnology Division, Defence Research and Development Establishment, Gwalior, India.
Sciex, A Division of DHR Holding India Pvt. Ltd., Gurgaon, India.
Lett Appl Microbiol. 2017 Mar;64(3):217-224. doi: 10.1111/lam.12706. Epub 2017 Jan 30.
Clostridium perfringens epsilon toxin, staphylococcal enterotoxin B and shiga toxin are implicated in a number of diseases and food-borne intoxications and are considered potential agents for bioterrorism and warfare. Artificially generated aerosol is the likely mode of delivery of these for nefarious uses, potentially capable of causing mass destruction to human and animal health by inhalation of toxic bioaerosol. Multiplex and unambiguous detection of these agents is of paramount importance for emergency response in a biothreat scenario and for food safety. Multiple-reaction monitoring (MRM) assay for simultaneous monitoring of the three toxins is reported here using reverse-phase high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry. Three different peptides with two fragment ions each were considered for quantification and confirmation. One of the three MRM transitions from each toxin, which exhibited the best sensitivity, was selected for multiplexing of the assay. Simulating a biothreat scenario wherein the bioaerosol is collected in 10 ml of buffer, the multiplex assay was tested with blind samples with one or more of the three toxins even in the presence of interfering Escherichia coli lysate proteins.
产气荚膜梭菌ε毒素、葡萄球菌肠毒素B和志贺毒素与多种疾病和食源性中毒有关，被认为是生物恐怖主义和战争的潜在制剂。人工制造的气溶胶可能是这些制剂用于邪恶目的的传播方式，通过吸入有毒生物气溶胶，有可能对人类和动物健康造成大规模破坏。对这些制剂进行多重且明确的检测对于生物威胁情况下的应急响应和食品安全至关重要。本文报道了使用反相高效液相色谱-电喷雾电离-串联质谱同时监测这三种毒素的多反应监测（MRM）分析方法。考虑了三种不同的肽，每种肽有两个碎片离子用于定量和确认。从每种毒素的三个MRM转换中选择一个灵敏度最佳的用于分析的多重检测。模拟生物威胁场景，即将生物气溶胶收集在10毫升缓冲液中，即使在存在干扰性大肠杆菌裂解物蛋白的情况下，也用含有三种毒素中一种或多种的盲样对多重分析进行了测试。
