Sugano Yohei, Sakata Kozue, Nakamura Kosuke, Noguchi Akio, Fukuda Nozomi, Suzuki Tomohiro, Kondo Kazunari
Hokkaido Institute of Public Health.
National Institute of Health Sciences.
Shokuhin Eiseigaku Zasshi. 2017;58(3):113-123. doi: 10.3358/shokueishi.58.113.
Omphalotus japonicus is a poisonous mushroom that grows in Japan. It can be mistaken for edible mushrooms (Shiitake, Hiratake and Mukitake), and if ingested, it causes food poisoning within 30 min to 1 hr. We established a rapid detection method using PCR-RFLP to identify O. japonicus by restriction digestion of the amplified ITS region. By using Sau96I, Bpu10I, SfcI or DrdI/HincII as a restriction enzyme, it was possible to rapidly identify and discriminate O. japonicus based on the fragment length. This study also provided a short PCR-RFLP system comprising amplification and digestion of a short 200-bp DNA fragment within the ITS region. The system could identify and discriminate O. japonicus after in vitro gastric digestion of native and heated mushroom samples as a model of food poisoning. In addition, a confirmatory assay using real-time PCR was developed to achieve more sensitive detection of O. japonicus.
日本发光脐菇是一种生长在日本的有毒蘑菇。它可能会被误认为是可食用蘑菇（香菇、平菇和滑菇），如果误食，会在30分钟到1小时内引发食物中毒。我们建立了一种利用PCR-RFLP的快速检测方法，通过对扩增的ITS区域进行限制性消化来鉴定日本发光脐菇。使用Sau96I、Bpu10I、SfcI或DrdI/HincII作为限制性内切酶，能够根据片段长度快速鉴定和区分日本发光脐菇。本研究还提供了一个简短的PCR-RFLP系统，该系统包括对ITS区域内一个200 bp的短DNA片段进行扩增和消化。作为食物中毒模型，该系统能够在对天然和加热的蘑菇样本进行体外胃消化后，鉴定和区分日本发光脐菇。此外，还开发了一种使用实时PCR的验证试验，以实现对日本发光脐菇更灵敏的检测。
