Rapid Identification Method of <i>Omphalotus japonicus</i> by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP)

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  • PCR-RFLPによるツキヨタケの迅速判別法
  • PCR-RFLP ニ ヨル ツキヨタケ ノ ジンソク ハンベツホウ

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Abstract

<p>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.</p>

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