AUTHOR=Long Pan , Jiang Zijuan , He Zhengmi , Chen Zuohong 

TITLE=Development of a loop-mediated isothermal amplification assay for the rapid detection of Russula subnigricans and Russula japonica

JOURNAL=Frontiers in Microbiology

VOLUME=13

YEAR=2022

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2022.918651

DOI=10.3389/fmicb.2022.918651

ISSN=1664-302X

ABSTRACT=<p><italic>Russula subnigricans</italic> is the only deadly species in the genus <italic>Russula</italic> with a mortality rate of more than 50%, and <italic>Russula japonica</italic> is the most common poisonous species, making rapid species identification in mushroom poisoning incidents extremely important. The main objective of this study was to develop a rapid, specific, sensitive, and simple loop-mediated isothermal amplification (LAMP) assay for the detection of <italic>R. subnigricans</italic> and <italic>R. japonica</italic>. Two sets of species-specific LAMP primers targeting internal transcribed spacer (ITS) regions were designed to identify <italic>R. subnigricans</italic> and <italic>R. japonica</italic>. The results demonstrated that while LAMP could specifically detect <italic>R. subnigricans</italic> and <italic>R. japonica</italic>, the polymerase chain reaction (PCR) could not distinguish <italic>R. subnigricans</italic> from <italic>Russula nigricans</italic>. In addition, the results demonstrated that, compared to electrophoresis-LAMP and real-time quantitative LAMP (RT-qLAMP), the detection sensitivity of HNB-LAMP (a mixture of LAMP with hydroxy naphthol blue (HNB) dye) for <italic>R. subnigricans</italic> could reach 0.5 pg/μl and was 100-fold higher than that of PCR. The LAMP reaction could be completed in 45 min, which is much faster than the conventional PCR. In the future, LAMP can be used a quick, specific, and sensitive detection tool in various fields.</p>