[Development and evaluation of a rapid PCR detection kit for Ophiocordyceps sinensis].
[Development and evaluation of a rapid PCR detection kit for Ophiocordyceps sinensis].
复制标题
DOI：
10.19540/j.cnki.cjcmm.20170217.011
复制
发表时间：
2017-03-01
期刊：
影响因子：
--
通讯作者：
Guo, Jin-Lin
中科院分区：
文献类型：
--
作者：
Hou, Fei-Xia;Cao, Jing;Guo, Jin-Lin
冬虫夏草是一种名贵中药材。由于资源紧缺、价格昂贵、市场需求量大，市场上存在大量的中华冬虫夏草掺假品。因此，有必要建立一种快速、有效的鉴别中华冬虫夏草的方法。本研究以中华水蛭种属特异性PCR为基础，通过成分优化开发了检测试剂盒，并对试剂盒的特异性、检出限、重复性和保质期进行了评价。结果表明，当冬虫夏草质量占该混合物的1/200以上时，即可成功检测。而且，只有中华水螅可以被扩增并在紫外光照射下发出明亮的绿色荧光。该试剂盒在37℃放置三天仍然有效，这表明该试剂盒在4℃保存一年稳定有效。该试剂盒在同批次不同操作条件下以及不同批次在相同操作条件下结果和准确度相同，说明该试剂盒重复性良好。利用该试剂盒可简单、快速、准确地区分中华药材及其掺伪品，为中药快速检测试剂盒的商业化奠定基础。
Ophiocordyceps sinensis is a valuable traditional Chinese medicine. Due to resource shortage, expensive price and huge market demand, there are many adulterants of O. sinensis in markets. Therefore, it is necessary to establish a rapid and effective method for distinguishing O. sinensis. Based on the species-specific PCR of O. sinensis, this study developed a detection kit by optimizing the components and evaluated the specificity, detection limit, repeatability and shelf life of the kit. The results showed that when the quality of O. sinensis accounted for more than 1/200 of that mixture, it could be detected successfully. Moreover, only O. sinensis could be amplified and glowed bright green fluorescence under ultraviolet light. The kit was still in effect when it was placed at 37 ℃ for three days, which indicated that it was stable and effective for one year stored in 4 ℃. The kit in the same batch under different operation conditions, and in different batch under the same operation conditions gave the same result and accuracy, which showed good repeatability of the kit. It is simple, rapid and accurate to distinguish O. sinensis from its adulterants using the kit, and lays the foundation for commercialization of traditional Chinese medicine fast detection kit.