目的 筛选龙胆Gentiana scabra Bunge、锁阳Cynomorium songaricum Rupr、天花粉Trichosanthes kirilowii Maxim.、菟丝子Cuscuta chinensis Lam.的DNA提取方法.方法 通过试剂盒法、SDS法、高盐低pH法、CTAB法以及改良PVP法,对4种中药材DNA进行提取.紫外分光光度、琼脂糖凝胶电泳和PCR扩增进行检测,SPSS20.0软件进行统计学分析.结果 改良PVP法所得4种中药材DNA的得率均较高.SDS法和改良PVP法提取的DNA条带较清晰,高盐低pH法提取者呈弥散状态,试剂盒法和CTAB法提取者几乎看不到条带.试剂盒法和改良PVP法提取的DNA可完全扩增ITS2和psbA-trnH序列,而其他3种方法未能完全扩增.结论 改良PVP法高效简单,最适合提取这4种中药材DNA.

Objective:To establish the simple and rapid examination system for the detection of Glycyrrhiza uralensis.Two detection methods,real-time PCR and LAMP,were developed for Glycyrrhiza uralensis detection,and then their specificity and sensitivity were compared.Methods:Extract the DNA of Glycyrrhiza uralensis and then amplify the special periods with designed primers.Process the red-time PCR and LAMP experiments to test the specificity of primers by the contrast of other species which were similar to Glycyrrhiza uralensis.Dilute the template concentration from 0.67 × 10 ng/μL to 0.67 × 10-5 ng/μL to make a contrast of the sensitivity between real-time PCR and LAMP.Results:Real-time PCR could amplify the special periods of Glycyrrhiza uralensis while other similar species was negative.The lowest concentration which real-time PCR could detect was 0.67 × 10-4 ng/μL.Similarly,the LAMP could detect the lowest concentration which was 0.67 × 10-4 ng/μL in 60 min which show the same sensitivity to the real-time PCR.Besides,the Lamp showed the specificity because other similar species couldn't amplify except Glycyrrhiza uralensis.It's obvious to observe green when put SYBR Green Ⅰ into the LAMP reaction product.Conclusion:The real-time PCR and the LAMP are all sensitive,accurate and specific in the detection However,the LAMP is more convenient and cost less time.What's more,the expensive equipment is not necessary for the LAMP.In a word,the LAMP is a better choice for the detection of Glycyrrhiza uralensis.