Abstract:
Mushrooms containing Amanita peptide toxins are the major cause of mushroom poisoning, and lead to approximately 90% of deaths. Phallotoxins are the fastest toxin causing poisoning among Amanita 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 Amanita phalloides 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.
摘要:
含有鹅膏肽毒素的蘑菇是蘑菇中毒的主要原因,并导致大约90%的死亡。在天牛肽毒素中,同调毒素是引起中毒最快的毒素。因此,构建一种高灵敏度的快速诊断毒蘑菇中毒的定量方法势在必行。在这项研究中,我们建立了一个高度敏感和自动化的磁珠(MB)为基础的化学发光免疫分析(CLIA),蘑菇中毒的快速诊断。在人血清中,同种异体毒素的检测限(LOD)为0.010ng/ml,在人尿中为0.009ng/ml。回收率为81.6%至95.6%,变异系数<12.9%。通过基于MB的自动化CLIA对天鹅膏样品的分析与HPLC-MS/MS的分析一致。基于MB的CLIA的优势,高灵敏度,重复性,和稳定性,是由于使用MB作为免疫载体,化学发光作为检测信号,和一个集成的设备,使整个过程自动化。因此,拟议的基于MB的自动化CLIA对于蘑菇中毒的早期和快速临床诊断是一个有希望的选择.