摘要
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 set 1 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.
引文
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