AUTHOR=He Zhengmi , Su Yuting , Li Sainan , Long Pan , Zhang Ping , Chen Zuohong 

TITLE=Development and Evaluation of Isothermal Amplification Methods for Rapid Detection of Lethal Amanita Species

JOURNAL=Frontiers in Microbiology

VOLUME=10

YEAR=2019

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2019.01523

DOI=10.3389/fmicb.2019.01523

ISSN=1664-302X

ABSTRACT=<p>In the present work, loop-mediated isothermal amplification (LAMP) and hyperbranched rolling circle amplification (HRCA) methods were developed to detect and distinguish different lethal <italic>Amanita</italic> species. Specific LAMP primers and HRCA padlock probes for species-specific identification and a set of universal LAMP primers for lethal <italic>Amanita</italic> species were designed and tested. The results indicated that the LAMP-based assay was able to discriminate introclade lethal <italic>Amanita</italic> species but was not able to discriminate intraclade species perfectly, while the HRCA-based assay could discriminate whether introclade or intraclade species. The universal LAMP primers were positive for 10 lethal species of <italic>Amanita</italic> section <italic>Phalloideae</italic> and negative for 16 species of <italic>Amanita</italic> outside section <italic>Phalloideae</italic>. The detection limits of LMAP and HRCA were 10 and 1 pg of genomic DNA per reaction, respectively. In conclusion, the two methods could be rapid, specific, sensitive and low-cost tools for the identification of lethal <italic>Amanita</italic> species.</p>