羚羊角塞特异性PCR鉴别方法研究

Molecular Authentication of Saigae Tataricae Cornu Tampon by Specific PCR

目的:为解决中药羚羊角塞与羚羊角相比因失去"通天眼"等传统鉴别特征而鉴别困难的问题,建立准确鉴别鉴定羚羊角塞的方法。方法:通过比对羚羊角塞基原动物赛加羚羊与其7种常见混伪品的细胞色素C氧化酶亚基I(Cytochrome coxidase I,COI)序列,根据差异位点设计出仅扩增赛加羚羊DNA的特异性鉴别引物。结果:进行PCR时,当退火温度为64℃,循环数为33个时,仅羚羊角塞正品扩增获得约300 bp大小的条带,混伪品无条带。结论:该方法可作为准确有效检测待检样品是否来源于赛加羚羊的方法,用于羚羊角类样品的DNA鉴别。

Objective： To solve the problem of authentication difficulty of saigae tataricae cornu tampon due to loss of traditional identify features such as ＂Tongtianyan＂,and to establish an accurate authentication method for saigae tataricae cornu tampon.Methods： Specific PCR primers for S. tatarica were designed based on cytochrome coxidase I（ COI） sequences and the annealing temperature of PCR reaction conditions was optimized,which was performed to authenticate saigae tataricae cornu tampon and its 7 adulterants. Results： Only saigae tataricae cornu tampon could obtain a ~ 300 bp specific band in electrophoresis while the adulterants did not when the temperature was 64 ℃ and 33 cycle number. Conclusion： Specific PCR could accurately and effectively authenticate the biological origin of sample is S. tatarica or not.