特异性PCR方法鉴别蛇胆汁及其伪品Identification of Fel Serpentis and its adulterants with specific PCR method
吴桂凡;张文娟;程显隆;谢培德;罗轶;石岩;魏锋;马双成;
WU Gui-fan;ZHANG Wen-juan;CHENG Xian-long;XIE Pei-de;LUO Yi;SHI Yan;WEI Feng;MA Shuang-cheng;Guangxi Institute for Food and Drug Control;National Institutes for Food and Drug Control;
摘要(Abstract):
建立一种简便准确的蛇胆汁药材DNA分子鉴别方法。测试并优化胆汁药材基因组DNA提取方法,并针对蛇类的COⅠ,Cytb,16S基因序列设计4对蛇类鉴别引物,根据特异性和扩增效率筛选最佳引物。使用其中1对引物Cytb1,20种蛇胆汁DNA均能扩增得到约400 bp的条带,同样条件下7种其他动物胆汁DNA均无扩增条带。该方法特异性强,准确度高,重复性好,为加强控制蛇胆汁药材质量提供了有效方法。
The study aims at developing a convenient and specific method for the identification of Fel Serpentis DNA. The methods of Fel Serpentis genomic DNA purification were tested and optimized,four pairs of specific primers for the amplification of COⅠ,Cyt b and 16 S were designed. Then the best pair of primers were selected according to the specificity and efficiency. The DNA fragment about 400 bp was amplified from 20 kinds of Fel Serpentis,whereas no DNA fragment was amplified from other animal samples under the same condition. This method is specific,accurate and reproducible,which provides a useful tool for the quality control of Fel Serpentis.
关键词(KeyWords):
蛇胆汁;鉴别;聚合酶链式反应(PCR);COⅠ;Cytb;16S
Fel Serpentis;PCR;COⅠ;Cytb;16S
基金项目(Foundation): 国家自然科学基金青年基金项目(31200673);; 国家“重大新药创制”科技重大专项(2014ZX09304307-001,2014ZX09304307-002)
作者(Author):
吴桂凡;张文娟;程显隆;谢培德;罗轶;石岩;魏锋;马双成;
WU Gui-fan;ZHANG Wen-juan;CHENG Xian-long;XIE Pei-de;LUO Yi;SHI Yan;WEI Feng;MA Shuang-cheng;Guangxi Institute for Food and Drug Control;National Institutes for Food and Drug Control;
Email:
DOI: 10.19540/j.cnki.cjcmm.2018.0067
参考文献(References):
- [1]李时珍.本草纲目.第4册[M].校点本.北京:人民卫生出版社,1981:2397.
- [2]中国药典.四部[S].2015:423.
- [3]薛漓.一种新的蛇胆掺假品的鉴别[J].中南药学,2003,1(5):305.
- [4]张穗.十四种蛇胆及其常见伪劣品的外观性状鉴别[J].蛇志,1992,4(2):55.
- [5]张小来.伪劣蛇胆的鉴别[J].现代应用药学,1990,7(3):20.
- [6]黄晓文,曾常青.眼镜蛇胆伪品鉴别[J].广东药学,2000,10(4):20.
- [7]刘向华,王义权,刘忠权,等.中药材蛇胆的DNA分子标记鉴定研究[J].药学学报,2001,36(3):229.
- [8]管淑玉,曾常青,刘广锋,等.中药蛇胆的鉴别及质量控制现状[J].广东药学院学报,2010,26(5):537.
- [9]廖婧,梁镇标,张亮,等.常见药用蛇类的DNA条形码研究[J].中国药学杂志,2013,48(15):1255.
- [10]黄勇,张月云,赵成坚,等.DNA条形码技术在常见中药材蛇类鉴别中的应用[J].中国中药杂志,2015,40(5):868.
- 吴桂凡
- 张文娟
- 程显隆
- 谢培德
- 罗轶
- 石岩
- 魏锋
- 马双成
WU Gui-fan- ZHANG Wen-juan
- CHENG Xian-long
- XIE Pei-de
- LUO Yi
- SHI Yan
- WEI Feng
- MA Shuang-cheng
- Guangxi Institute for Food and Drug Control
- National Institutes for Food and Drug Control
- 吴桂凡
- 张文娟
- 程显隆
- 谢培德
- 罗轶
- 石岩
- 魏锋
- 马双成
WU Gui-fan- ZHANG Wen-juan
- CHENG Xian-long
- XIE Pei-de
- LUO Yi
- SHI Yan
- WEI Feng
- MA Shuang-cheng
- Guangxi Institute for Food and Drug Control
- National Institutes for Food and Drug Control