Abstract：
Amanita exitialis, a deadly mushroom found in eastern Asia, causes the highest death rates among all poisonous mushrooms in China. The aim of the present study was to develop an efficient, accurate, and user-friendly PCR-based method for identifying A. exitialis that could facilitate the prevention, diagnosis, and treatment of associated food poisoning. A. exitialis-specific primers and probes were designed based on the internal transcribed spacer region variations of 27 mushroom species. Specificity was confirmed using conventional and real-time PCR for 23 non-target mushroom species, including morphologically similar and closely related species. Compared to conventional PCR, real-time PCR was more sensitive (detectable DNA concentration: 1.36 × 10-2 ng/μL vs. 1.36 × 10-3) and efficient (analysis time: 1 h vs. 40 min). Furthermore, the real-time PCR results could be immediately visualized using amplification curve analysis. The results present two robust PCR-based methods for A. exitialis identification that can facilitate food safety.
摘要：
白天牛，一种在东亚发现的致命蘑菇，在中国所有毒蘑菇中死亡率最高。本研究的目的是开发一种有效的，准确,和用户友好的基于PCR的方法，用于鉴定可以促进预防的近侧索杆菌，诊断,以及相关食物中毒的治疗。根据27个蘑菇物种的内部转录间隔区变异设计了天然特异性引物和探针。使用常规和实时PCR对23种非目标蘑菇物种进行了特异性确认，包括形态相似和密切相关的物种。与常规PCR相比,实时PCR更敏感（可检测的DNA浓度：1.36×10-2ng/μL与1.36×10-3）和有效（分析时间：1hvs.40分钟）。此外，实时PCR结果可以使用扩增曲线分析立即可视化。结果提供了两种基于PCR的可靠方法，可用于促进食品安全。
