Identification of Gyromitra infula: A Rapid and Visual Method Based on Loop-Mediated Isothermal Amplification.

Xiaomei Xie,Renhe Duan,Yuan Zheng,Yuguang Fan,Enjing Tian,Bu Li,Chonghua Gao

doi:10.3389/fmicb.2022.842178

Abstract

With mushroom poisoning emerging as one of the most serious food safety problems worldwide, a rapid identification method of poisonous mushrooms is urgently required to investigate the source of poisoning. Gyromitra infula, a kind of poisonous mushroom, contains gyromitrin toxin, which causes epileptogenic neurotoxicity and hemolytic disease. This study aimed to establish a rapid and visual method of G. infula identification based on loop-mediated isothermal amplification (LAMP). A set of specific LAMP primers was designed, and its specificity in G. infula was confirmed against various mushroom species, including its closely related species and other macrofungi. The sensitivity assay showed that the minimum concentration of genomic DNA detected by LAMP was 1 ng/μl. The method’s applicability was conducted by preparing mushroom samples that were boiled and digested in artificial gastric juice. The results showed that the content as low as 1% G. infula can be successfully detected. This method can be completed within 90 min, and the reaction results can be directly observed by the naked eyes. Hence, the identification method of G. infula established based on LAMP in this study is accurate, rapid, sensitive, and low-cost, which is required for clinical treatment or forensic analysis when mushroom poisoning occurs.

Highlights

Mushrooms are widely distributed globally, some of which have high nutritional value and medicinal efficacy (Lei et al, 2020), but some of which are poisonous
We carried out three replicates and each replicate showed the same result. This further proved that the loop-mediated isothermal amplification (LAMP) primer set in this experiment only amplified the DNA of G. infula
To further validate the specificity of primer set I, the control primer set II in Table 2 was used for the LAMP control experiment of G. infula and closely related species

Summary

Introduction

Mushrooms are widely distributed globally, some of which have high nutritional value and medicinal efficacy (Lei et al, 2020), but some of which are poisonous. There are more than 4,000 mushrooms species in China, among which 966–1,020 are edible (Dai et al, 2010; Wu et al, 2019), 692 are medicinal (Wu et al, 2019), and 435–480 are poisonous (Bau et al, 2014; Wu et al, 2019). Mushroom poisoning mostly occurs from June to October annually. Mushroom poisoning is mostly common in Southwest China and Central. Poisonous mushrooms can cause poisoning, they play a certain role in some fields, such as medicine and insecticides, due to their biologically active compounds (Wu et al, 2019; Sheikha, 2021)

Objectives

Methods

Results

Discussion

Conclusion