Rapid identification of Lepiota brunneoincarnata in China with loop-mediated isothermal amplification

IntroductionMany edible fungi and poisonous mushrooms share similar morphologic characteristics, making them difficult to distinguish with the naked eye. Lepiota brunneoincarnata is a major lethal toxic mushroom widely distributed in China and responsible for frequent poisoning incidents across the country.MethodsThe present study developed a loop-mediated isothermal amplification (LAMP) reaction with a primer set designed from the translation elongation factor 1-a (tef1) sequence of L. Brunneoincarnata.ResultsA set of specific LAMP primers were designed for the specific detection of L. brunneoincarnata, and the assay showed no amplification from 63 non-target fungal specimens. In this experiment, the limit of detection for DNA concentration was 100 pg/mL, and the limit of detection for spore concentration was 10 spores/mL. The method also successfully detected L. brunneoincarnata in boiled and digested mixed mushroom samples, and the whole detection procedure can be completed within 60 minutes at 63 °C.DiscussionThe study is suitable for rapid identification of L. brunneoincarnata in China, and the method has potential for further optimization toward field application.