著者
Sayuri TSURUDA Kouichi AKAKI Hiroshi HIWAKI Akira SUZUKI Hiroshi AKIYAMA
出版者
(社)日本農芸化学会
雑誌
Bioscience, Biotechnology, and Biochemistry (ISSN:09168451)
巻号頁・発行日
vol.76, no.7, pp.1343-1349, 2012-07-23 (Released:2012-07-23)
参考文献数
33
被引用文献数
8

A rapid multiplex real-time PCR assay was developed to achieve highly specific, simultaneous detection of two kinds of mushrooms, Omphalotus guepiniformis and Lentinula edodes. Primers and TaqMan minor groove binder probes were designed according to the internal transcribed spacers 1-5.8S region of rDNA and evaluated by the specificity for fruiting bodies of 17 O. guepiniformis, 16 L. edodes and samples from 57 other species. DNA extracts of all the target species had positive signals with no cross-reaction, the limit of detection being 0.00025 ng of DNA. Threshold cycle (Ct) values for raw and processed fruiting bodies and for fruiting bodies (1% (w/w)) mixed with foodstuffs or artificial gastric juice contents ranged from 17.16 to 26.60 for both examined species. This new assay proved specific to the target species, highly sensitive, and applicable to processed food samples and gastric juice contents, making it useful for rapidly identifying O. guepiniformis and L. edodes.