PCR-RFLP定量检测川贝母真伪的研究

Authentication of Fritillariae cirrhosae bulbus by PCR-RFLP

建立了一个适合中药材川贝母的聚合酶链式反应-限制性内切酶(PCR-RFLP)的分子鉴定方法。以提取的DNA为模板进行PCR扩增和酶切反应,真品川贝母扩增产物含有Sam I酶切位点,在100~250 bp之间存在明确的2条酶切条带,而伪品贝母扩增序列中没有Sam I酶切位点不能被切出。此外,还基于PCR-RFLP技术针对川贝母真伪性进行了相对定量分析研究。结果表明,含量10%及以上的真品川贝母能被稳定检测出。

In this study,a molecular identification method of polymerase chain reaction restriction endonuclease(PCR-RFLP)was developed for identification of Fritillariae cirrhosae bulbus.The extracted DNA was used as the template for PCR amplification,and the amplification products were digested with endonuclease.Because Fritillariae cirrhosae bulbus has Sam I cut site,2 distinct bands between 100 bp to 250 bp could be observed.However,the pseudo Fritillaria could not be cut.Based on the PCR-RFLP technique,the relative quantitative analysis standardization of Fritillariae cirrhosae bulbus was also studied.The results showed that products containing 10%or more authentic Fritillariae cirrhosae bulbus could be detected.