Zhu Jianyu, Dou Leina, Shao Shibei, Kou Jiaqian, Yu Xuezhi, Wen Kai, Wang Zhanhui, Yu Wenbo
Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety Beijing Laboratory for Food Quality and Safety, College of Veterinary Medicine, China Agricultural University, Beijing, China.
School of Basic Medicine, Beihua University, Jilin, China.
Front Chem. 2021 Dec 23;9:813219. doi: 10.3389/fchem.2021.813219. eCollection 2021.
Mushrooms containing peptide toxins are the major cause of mushroom poisoning, and lead to approximately 90% of deaths. Phallotoxins are the fastest toxin causing poisoning among peptide toxins. Thus, it is imperative to construct a highly sensitive quantification method for the rapid diagnosis of mushroom poisoning. In this study, we established a highly sensitive and automated magnetic bead (MB)-based chemiluminescence immunoassay (CLIA) for the early, rapid diagnosis of mushroom poisoning. The limits of detection (LODs) for phallotoxins were 0.010 ng/ml in human serum and 0.009 ng/ml in human urine. Recoveries ranged from 81.6 to 95.6% with a coefficient of variation <12.9%. Analysis of samples by the automated MB-based CLIA was in accordance with that of HPLC-MS/MS. The advantages the MB-based CLIA, high sensitivity, repeatability, and stability, were due to the use of MBs as immune carriers, chemiluminescence as a detection signal, and an integrated device to automate the whole process. Therefore, the proposed automated MB-based CLIA is a promising option for the early and rapid clinical diagnosis of mushroom poisoning.
含有肽类毒素的蘑菇是蘑菇中毒的主要原因，约导致90%的死亡。鬼笔毒素是肽类毒素中导致中毒最快的毒素。因此，构建一种用于蘑菇中毒快速诊断的高灵敏度定量方法势在必行。在本研究中，我们建立了一种基于磁珠（MB）的高灵敏度自动化化学发光免疫分析（CLIA）方法，用于蘑菇中毒的早期快速诊断。鬼笔毒素在人血清中的检测限（LOD）为0.010 ng/ml，在人尿中的检测限为0.009 ng/ml。回收率在81.6%至95.6%之间，变异系数<12.9%。基于自动化MB的CLIA对样品的分析结果与HPLC-MS/MS的分析结果一致。基于MB的CLIA具有高灵敏度、重复性和稳定性等优点，这归因于使用MB作为免疫载体、化学发光作为检测信号以及使用集成设备实现整个过程的自动化。因此，所提出的基于自动化MB的CLIA是蘑菇中毒早期快速临床诊断的一个有前景的选择。
