University of Applied Sciences, HES-SO Valais//Wallis, Institute of Life Technologies, Sion, Switzerland.
University of Montreal, Institute for Research in Immunology and Cancer, Montreal, Quebec, Canada.
Sci Rep. 2019 Apr 9;9(1):5814. doi: 10.1038/s41598-019-42167-0.
Cereulide is a toxic cyclic depsipeptide produced by certain strains of Bacillus cereus found in soil and food products. While some harmless strains of Bacillus are used as probiotic, others can cause nausea and vomiting, and represent an important food safety concern. Current detection methods are time consuming and do not necessarily detect toxic cereulide. Here, we developed a rapid protocol using Matrix Assisted Laser Desorption/Ionization-Time of Flight (MALDI-TOF) mass spectrometry that detects the toxin originating from a colony smear of B. cereus. The distinct molecular feature of the toxin peak at m/z 1,191 was clearly identified from bacterial extracts with a limit of detection (LOD) of 30 ng/mL. Final optimisation of the sample preparation was based on cereulide chelating cations to produce the alkali adduct [M + K] without the use of a MALDI matrix, and provided a 1,000-fold improvement of LOD with 30 pg/mL of cereulide. We evaluated the application of this method for the detection of cereulide in rice, milk, and different ready-to-eat meals. The proposed protocol is quick, easy and provides an improvement over conventional methods for the detection of B. cereus toxin.
脑苷脂是一种由土壤和食品产品中某些蜡样芽孢杆菌菌株产生的有毒环二肽。虽然某些无害的芽孢杆菌菌株被用作益生菌,但其他菌株可能会引起恶心和呕吐,这是一个重要的食品安全问题。目前的检测方法既耗时又不一定能检测到有毒的脑苷脂。在这里,我们开发了一种使用基质辅助激光解吸/电离飞行时间(MALDI-TOF)质谱的快速检测方法,用于检测源于蜡样芽孢杆菌菌落涂片的毒素。通过细菌提取物中 m/z 1191 处的毒素峰的明显分子特征,该方法可以明确检测到 30ng/mL 的检测限(LOD)。最终优化了样品制备方法,即通过螯合阳离子将脑苷脂转化为碱加合物 [M+K],而无需使用 MALDI 基质,从而使脑苷脂的 LOD 提高了 1000 倍,达到 30pg/mL。我们评估了该方法在检测大米、牛奶和不同即食餐中的脑苷脂方面的应用。该方法快速、简单,并且比传统方法在检测蜡样芽孢杆菌毒素方面有了改进。