Multiplex Real-Time PCR Assay for Simultaneous Detection of Omphalotus guepiniformis and Lentinula edodes

被引：13

作者：

Tsuruda, Sayuri ^{[1
]}

Akaki, Kouichi ^{[1
]}

Hiwaki, Hiroshi ^{[1
]}

Suzuki, Akira ^{[2
]}

Akiyama, Hiroshi ^{[3
]}

机构：

[1] Fukuoka City Inst Hyg & Environm, Chuo Ku, Fukuoka 8100065, Japan

[2] Chiba Univ, Fac Educ, Inage Ku, Chiba 2638522, Japan

[3] Natl Inst Hlth Sci, Setagaya Ku, Tokyo 1588501, Japan

来源：

BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY | 2012年 / 76卷 / 07期

关键词：

species identification; multiplex real-time PCR; Omphalotus guepiniformis; Lentinula edodes; poisonous mushroom; POLYMERASE-CHAIN-REACTION; ILLUDIN-S; POISONOUS MUSHROOMS; MASS-SPECTROMETRY; QUANTITATIVE PCR; FOOD; IDENTIFICATION; MAIZE; CHROMATOGRAPHY; DEGRADATION;

D O I：

10.1271/bbb.120090

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：1343 / 1349

页数：7

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