Development of Loop-Mediated Isothermal Amplification (LAMP) Methods for Rapid Identification of Nephrotoxic Amanita...

Mushroom poisoning remains the leading cause of foodborne morbidity and mortality in China. The mushrooms that cause acute kidney injury (AKI) belong to the species of Amanita genus section Roanokenses. These species exhibit highly similar morphological characteristics, which is a challenge for their identification for non-specialists. Although conventional DNA barcoding identification is reliable, it is time-consuming for rapid diagnosis. In this study, we developed a loop-mediated isothermal amplification (LAMP) method based on the internal transcribed spacer (ITS) region to enable rapid identification of five species within A. sect. Roanokenses which can cause AKI. The optimized LAMP system was rigorously evaluated for specificity and sensitivity, with comparative analysis performed against conventional PCR. The results demonstrated that the established LAMP method achieved visual detection without specialized equipment, exhibiting high specificity and a detection limit of 10 pg per reaction. Critically, this approach can reduce identification time to approximately 1 h and provides sensitivity 100 times greater than that of the conventional PCR protocol.