An Automated and Highly Sensitive Chemiluminescence Immunoassay for Diagnosing Mushroom Poisoning

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.

Molecular Recognition Mechanism of an Anti-Amatoxins mAb and Its Application in Centrifugal Disk-Based Immunoassay

“…IMBs as carriers in immunoassay were obtained by coupling MBs-Avi and biotin ester-labeled coating antigen (α-AMA-OVA−Bio) using the biotin-avidin system. 37 With the help of IMBs and a magnetic field provided by the MI600 controller, multistep immunoassay operations including sample adding, washing, and signal generation can be automatically achieved on a centrifugal microfluidic platform for rapid detection of amatoxins. As shown in Figure 3A,B, the CD-like chip was composed of 12 units, and quantitative detection of 12 samples could be performed simultaneously.…”

“…Centrifugal microfluidics is a microfluidic technology that manipulates liquid in disks using centrifugal forces. IMBs as carriers in immunoassay were obtained by coupling MBs-Avi and biotin ester-labeled coating antigen (α-AMA-OVA–Bio) using the biotin-avidin system . With the help of IMBs and a magnetic field provided by the MI600 controller, multistep immunoassay operations including sample adding, washing, and signal generation can be automatically achieved on a centrifugal microfluidic platform for rapid detection of amatoxins.…”

Mushroom poisoning outbreaks in Guizhou Province, China: a prediction study using SARIMA and Prophet models

“…The Prophet and SARIMA models predicted an increase in exposures, cases, and hospitalizations, with the peak reporting period continuing to be the rainy season from June to September. Fortunately, there is a definite decreasing trend in the total number of fatalities, whereas seasonal patterns indicate that October is the month with the highest number of fatalities owing to the deadly fungal species Amanita exitialis 33 , 34 . The probability of death from mushroom poisoning is highly dependent on the type of mushroom; for example, highly toxic mushrooms of the Amanita exitialis cause acute liver damage and have a much higher mortality rate than other generally toxic mushrooms.…”