Long Pan, Jiang Zijuan, He Zhengmi, Chen Zuohong
College of Life Science, Hunan Normal University, Changsha, China.
Front Microbiol. 2022 Aug 23;13:918651. doi: 10.3389/fmicb.2022.918651. eCollection 2022.
is the only deadly species in the genus with a mortality rate of more than 50%, and 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 and . Two sets of species-specific LAMP primers targeting internal transcribed spacer (ITS) regions were designed to identify and . The results demonstrated that while LAMP could specifically detect and , the polymerase chain reaction (PCR) could not distinguish from . 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 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.
是该属中唯一死亡率超过50%的致命物种，并且是最常见的有毒物种，这使得在蘑菇中毒事件中快速进行物种鉴定极为重要。本研究的主要目的是开发一种用于检测和的快速、特异、灵敏且简单的环介导等温扩增（LAMP）检测方法。设计了两组针对内部转录间隔区（ITS）区域的物种特异性LAMP引物来鉴定和。结果表明，虽然LAMP能够特异性检测和，但聚合酶链反应（PCR）无法区分和。此外，结果表明，与电泳-LAMP和实时定量LAMP（RT-qLAMP）相比，HNB-LAMP（LAMP与羟基萘酚蓝（HNB）染料的混合物）对的检测灵敏度可达0.5 pg/μl，比PCR高100倍。LAMP反应可在45分钟内完成，比传统PCR快得多。未来，LAMP可作为各个领域快速、特异且灵敏的检测工具。
