基于LAMP的中药材川贝母真伪鉴别方法的建立

目的：基于荧光-环状等温扩增技术，建立川贝母快速检测方法，实现中药材川贝母的荧光可视化快速鉴定。方法：依据川贝母物种特异性序列设计5套LAMP引物，并测试方法特异性和灵敏度。结果：引物Set1扩增效率最高；使用该引物建立的川贝母LAMP检测方法仅能够在6种川贝母基源药材中扩增出扩增曲线和荧光变化，在伊贝母、浙贝母、平贝母等7种常见混淆品中无扩增产物，方法特异性良好；方法检测灵敏度为0.1%。结论：本研究建立的基于LAMP的川贝母鉴定方法具有较好的特异性和灵敏度，其快速、操作简单、易观察的优点，填补了中药材川贝母真伪快速筛查的技术空白。

Objective: To establish rapid visual identification method for Fritillaria cirrhosa based on loop-mediated isothermal amplification. Methods: According to the DNA sequence of the target,five sets of primers were designed and screened by amplification efficiency. Specificity and sensitivity of the method was validated. Results: The LAMP detection method established by using primer set1 showed best performance on amplification efficiency, specificity and sensitivity. Turbidity and fluorescence changes could be observed in 6 kinds of Fritillariae, and there was no amplification product in 7 common adulterants. The sensitivity of the LAMP method was 0.1%. Conclusion: The proposed LAMP method is specific, sensitive, accurate and simple and easy to observe, which fills the gap for the authentication of Fritillaria cirrhosa.

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