Rapid identification of Aconitum plants based on loop-mediated isothermal amplification assay

Abstract

ObjectiveAconitum plants (Ranunculaceae) exhibit toxicity, and accidental ingestion of the plants has been reported in Japan. Identifying the cause of poisoning is important for emergency medical treatment, and a rapid and simple detection technique is required for the identification of poisoning cause. In the present study, we developed a rapid and simple method for detecting Aconitum plant DNA using a loop-mediated isothermal amplification (LAMP) assay.ResultsSpecific LAMP primers for Aconitum plants were designed based on the trnL–trnF intergenic spacer region. Using the LAMP primers, the LAMP assay included an initiation reaction of 10\xa0min followed by amplification for 20\xa0min at the isothermal reaction temperature of 65\xa0°C. The LAMP reaction was demonstrated to be specific and highly sensitive to Aconitum plants, given that the assay can be used for 1\xa0pg of purified DNA. Using raw extracted DNA as template, the entire detection procedure from DNA extraction to final detection required only 30\xa0min. Moreover, the protocol identified samples containing approximately 5\xa0mg of Aconitum plants cooked and digested with artificial gastric juice. The currently proposed protocol exhibits good potential as a screening method of Aconitum plant poisoning for emergency medical care.