活血止痛胶囊中土鳖虫的分子鉴定研究

Molecular Identification of the Cockroach Eupolyphaga sinensis in a Compound Chinese Medicine,Huoxuezhitong Capsule

目的建立活血止痛胶囊中土鳖虫的一种分子鉴定方法。方法采用蛋白酶K法结合吸附柱法提取活血止痛胶囊中土鳖虫基因组DNA,使用蜚蠊目昆虫通用引物扩增其线粒体16S rRNA片段并测序。结果土鳖虫能扩增出475 bp左右的片段,本实验成功从自制活血止痛胶囊中扩增出该片段。结论该方法灵敏度高,可靠性强,可以做为活血止痛胶囊中土鳖虫的一种鉴定方法。

OBJECTIVE To establish a molecular determination method of the cockroach Eupolyphaga sinensis in a compound Chinese medicine,Huoxuezhitong Capsule.METHODS The proteinase K conjugation adsorption column method was used to extract and purify genomic DNA.Using the universal primer of Blattaria,16S rRNA was amplified and sequenced.RESULTS A fragment about 475 bp was obtained from Eupolyphaga sinensis.The self-made Huoxuezhitong capsule by different processing methods could be amplified successfully.Eupolyphaga sinensis and Huoxuezhitong capsule purchased from the market could not be amplified.CONCLUSION The method is reliable,and can be used for quality control of Huoxuezhitong capsule.