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基于Taqman-MGB探针的亚稀褶红菇实时荧光定量PCR检测方法
引用本文:章轶哲,李宏锡,张凯平,孙承业,张宏顺,卢俊伽,何仟,李海蛟.基于Taqman-MGB探针的亚稀褶红菇实时荧光定量PCR检测方法[J].菌物学报,2020,39(5):937-943.
作者姓名:章轶哲  李宏锡  张凯平  孙承业  张宏顺  卢俊伽  何仟  李海蛟
作者单位:1. 中国疾病预防控制中心职业卫生与中毒控制所 北京 1000502. 广东省茂名市野生动物救护研究中心 广东 茂名 525000
摘    要:本研究建立了一种基于Taqman-MGB探针的亚稀褶红菇Russula subnigricans实时荧光定量PCR检测方法。根据亚稀褶红菇与其近似种的内转录间隔区(internal transcribed spacers,ITS)序列差异,设计合成1对引物和1条特异性Taqman-MGB探针,并用常见有毒红菇种类进行验证。结果显示,引物特异性良好,仅亚稀褶红菇出现荧光信号,完成整个检测过程只需2h。该法能够为毒蘑菇中毒的快速检测提供技术支持。

关 键 词:蘑菇中毒  亚稀褶红菇  快速检测  实时荧光定量PCR  Taqman-MGB探针  
收稿时间:2019-09-04

A method of real-time fluorescent quantitative PCR for detection of Russula subnigricans based on Taqman-MGB probe
Authors:Yi-Zhe ZHANG  Hong-Xi LI  Kai-Ping ZHANG  Cheng-Ye SUN  Hong-Shun ZHANG  Jun-Jia LU  Qian HE  Hai-Jiao LI
Institution:1. National Institute of Occupational Health and Poison Control, Chinese Center for Disease Control and Prevention, Beijing 100050, China2. Rescue and Research Center for Wild Animals of Maoming, Maoming, Guangdong 525000, China
Abstract:A method for detection of Russula subnigricans was established based on Taqman-MGB probe of real-time fluorescent quantitative PCR. A pair of primers and a Taqman-MGB probe were designed and synthesized according to the ITS sequences of R. subnigricans and its related species, and the common toxic Russula species were used for verification. The results showed that the primers had good specificity, and only R. subnigricans exhibited fluorescence signal. It just took 2h to complete the whole detection process. It was indicated that this method could provide a technical support for rapid detection of toxic mushroom poisoning.
Keywords:mushroom poisoning  Russula subnigricans  rapid detection  real-time fluorescent quantitative PCR  Taqman-MGB probe  
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