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A Rapid colorimetric assay for field-portable authentication of Amanita sepiacea using the beta-tubulin gene sequence

Jie Gao, Nan Wang, Ruibin Xie, Juan Zhang, Enjing Tian, Hongjing Wang, Jianxin Tan, Ailiang Chen
Journal of food composition and analysis 2022 v.105 pp. 104169
Amanita, DNA, colorimetry, food composition, genes, loop-mediated isothermal amplification, mushrooms, nontarget organisms, nucleotide sequences, rapid methods, toxicity, tubulin
Globally, mushroom poisoning is becoming one of the most serious food safety-related problems. To investigate the source of mushroom poisoning, a rapid method for identifying toxic mushrooms is urgently required. This study aimed to establish a colorimetric method based on loop-mediated isothermal amplification (LAMP) for rapid detection of Amanita sepiacea based on the analysis of the beta-tubulin gene. Specific LAMP primers for species specificity identification were designed. The primers designed by the colorimetric method did not cross-react with 41 non-target species mushrooms, and 1 ng of A. sepiacea DNA was detected. The detection method established herein can be used to identify A. sepiacea when present at concentrations as low as 1% in mixed fresh, cooked, or fried mushroom samples. The whole detection process only takes 30 min and requires heating and no complex instrumental help. Therefore, this method can be used in the field detection of poisonous mushrooms.