Species identification of white false hellebore (Veratrum album subsp. oxysepalum) by loop-mediated isothermal amplification (LAMP)

Abstract

Purpose: Food poisoning is frequently caused by the accidental ingestion of toxic plants with strong morphological similarities to edible plants. The plant, false helleborine (Veratrum album), is commonly involved in such incidents. Cases of poisoning by toxic plants require rapid and accurate identification for appropriate medical treatment or forensic investigation in the event of death. However, morphological examinations require experience in systematic botany, and chemical analysis of natural compounds can be challenging. This study aimed to develop a sensitive, rapid, and specific method for identifying V. album and V. stamineum. Methods: We developed and optimized a loop-mediated isothermal amplification (LAMP) detection method. Total genomic DNA was extracted from approximately 10 mg (dry weight) of leaf from 26 specimens comprising the target plants, allied plants, and other causative plants of poisoning. These samples were analyzed the LAMP assay, and mixed samples, cooked samples, and simulated gastric contents were also tested. Results: The limit of detection of our LAMP assay was 0.08 ng of target DNA, and the assay was specific to V. album and V. stamineum. The LAMP assay could be performed in approximately 30 min. V. album was also successfully identified in mixed samples, cooked samples, and simulated gastric contents. In addition, the LAMP assay signal could be effectively assessed with the naked eye. Conclusions: The present LAMP assay is a highly sensitive, rapid, and specific method for identifying V. album and V. stamineum.