摘 要 :目的:寻找天花粉类药材快捷准确的分子鉴别方法。方法:在天花粉ITS保守区域设计3个20 mer左右单引物,对19批天花粉商品及其9种伪品进行PCR扩增。结果:引物TkS1-64得到天花粉及其伪品的DNA多态性图谱,其中560 bp和960 bp条带为天花粉的特征鉴别条带。结论:引物TkS1-64具有稳定性好、重复性高的优点,可以作为天花粉类药材分子鉴别引物。此方法定名为锚定引物扩增多态性DNA(anchored primer amplification polymorphism DNA,简称APAPD),在中药材鉴别中具有推广应用的价值。
Abstract:Objective: To explore a new molecular method to authenticate Radix Trichosanthis. Method: Three 20 mer primers based on the ITS sequence was designed.The PCR reaction system was optimized and applied to nineteen different sources of Radix Trichosanthis and nine adulterants and substitutes.Result: Polymorphic map of Radix Trichosanthis and its adulterants was obtained from primer TKS1-64. 560 bp and 960 bp bands were authentic markers for Radix Trichosanthis. Conclusion: Primer TKS1-64F possesses the advantages of good stability and reproducibility. This new method is named as anchored primer amplification polymorphism DNA(APAPD). It was a potential method to used in molecular identification of other meteria medica.