Objective Analyzing on-site disposal process of a mushroom poisoning in Wuhan. Identifying the toxin in the mushrooms and providing references for similar poisoning incidents. Methods Epidemiological investigation, clinical treatment data and suspicious poisonous mushroom sample were collected. Mushroom sample was identified by molecular analysis. Results All patients ate different quantity of fried mushroom. The early clinical symptom were mainly gastrointestinal symptoms, and the liver, kidney and coagulopathy were damaged in the later stage. The mushroom samples were identified as Amanita rimosa. Conclusion According to epidemiological,clinical,morphological and molecular biological evidence,this incident was confirmed to be a mushroom poisoning incident caused by Amanita rimosa.