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冬虫夏草为我国传统名贵中药材。资源短缺、价格昂贵和巨大的市场需求导致市售冬虫夏草掺伪掺假现象严重,亟需建立快速有效的鉴别方法。该研究基于实验前期已建立的冬虫夏草快速鉴定SS-PCR方法,按照体外诊断试剂评价标准开发试剂盒,并对试剂盒的特异性、检测限、重复性及保存期4项性能进行了评价。结果显示冬虫夏草质量占混合物质量的1/200(质量分数0.5%以上)以上时可被成功检测。试剂盒对冬虫夏草及其伪品的检测结果显示,仅有冬虫夏草可被成功扩增,并在紫外下显示绿色荧光。该试剂盒37℃温育7 d仍有效,说明4℃保存其在1年内稳定有效。同一批次试剂盒在不同条件下,不同批次试剂盒在相同条件下,其检测结果均一致且准确,表明试剂盒具有良好的批内及批间重复性。将该试剂盒用于市售冬虫夏草及其混伪品的检测,操作简单快速,结果准确可靠,为下一步中药快检试剂盒的商品化奠定了基础。
Cordyceps sinensis for our traditional precious Chinese herbal medicines. Shortage of resources, high prices and huge market demand led to the adulteration of commercially available Cordyceps sinensis serious phenomenon, the urgent need to establish a rapid and effective identification methods. The Rapid Determination SS-PCR method is based on preliminary experiments Cordyceps has been established in accordance with the evaluation criteria developed in vitro diagnostic reagent kit, and specific kit, detection limit, reproducibility and shelf life of four properties were evaluated. The results showed that the quality of Cordyceps mixture of the quality of 1/200 (mass fraction of 0.5% or more) can be successfully detected. The test results of the kit on Cordyceps sinensis and its counterfeit products showed that only Cordyceps sinensis could be successfully amplified and showed green fluorescence under ultraviolet light. The kit incubated at 37 ℃ for 7 days is still valid, indicating that 4 ℃ stored in a stable and effective within 1 year. Under the same conditions of different batches of kits in the same batch of kits, the test results of different batches of kits are consistent and accurate under the same conditions, indicating that kits have good intra-batch and inter-batch repeatability. The kit for the detection of commercially available Cordyceps and its adulterants, simple and fast operation, the results are accurate and reliable, for the next step of commercialization of Chinese medicine rapid test kit to lay the foundation.