中国中药杂志

2018, v.43(05) 945-951

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牛膝及川牛膝配方颗粒位点特异性PCR鉴别研究
Allele-specific PCR to identification of Achyranthis Bidentatae Radix and Cyathulae Radix formula granules

孟虎彪;袁媛;刘富艳;蒋超;金艳;赵玉洋;
MENG Hu-biao;YUAN Yuan;LIU Fu-yan;JIANG Chao;JIN Yan;ZHAO Yu-yang;State Key Laboratory of Dao-di Herbs Breeding Base,National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences;

摘要(Abstract):

为建立一种稳定准确鉴定牛膝和川牛膝配方颗粒的方法。该文使用位点特异性PCR技术,根据牛膝和川牛膝ITS序列的SNP位点设计特异性鉴别引物,经过优化DNA提取方法后,使用牛膝特异性引物进行扩增,牛膝基原植物和牛膝配方颗粒均可扩增出187 bp特异性条带,川牛膝及混伪品无条带;使用川牛膝特异性引物进行扩增,川牛膝基原植物和川牛膝配方颗粒均可扩增出162 bp特异性条带。并利用所建立的方法对不同生产企业的牛膝和川牛膝配方颗粒进行鉴别,可以充分弥补传统鉴别方法的局限性。
To establish a robust and accuracy molecular method to identify Achyranthis Bidentatae Radix and Cyathulae Radix formula granules. ITS sequences of Achyranthes bidentata and Cyathula officinalis were aligned,specific SNPs(single nucleotide polymorphisms) were excavated,specific primers were designed and allele-specific PCR method was established. The genomic DNA was successfully extracted from the herbal medicine and its formula granules by using an improved CTAB(cetyltrimethyl ammonium bromide) method and then performed PCR with the designed primers. The 187 bp specific band could be amplified only in the presentation of C. officinalis and its granules when use of C. officinalis specific primers,whereas the 162 bp band could be amplified only in the presentation of A. bidentata and its granules when use of A. bidentata specific primers. This method was also successfully applied in the identification of commercial formula granules.

关键词(KeyWords): 牛膝;川牛膝;配方颗粒;位点特异性PCR;分子鉴定
Achyranthes bidentata;Cyathula officinalis;formula granule;allele-specific PCR;molecular authentication

Abstract:

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基金项目(Foundation): 中医药行业科研专项(201407003);; 中央级公益性科研院所基本科研业务费专项资金项目(ZZ10-008)

作者(Author): 孟虎彪;袁媛;刘富艳;蒋超;金艳;赵玉洋;
MENG Hu-biao;YUAN Yuan;LIU Fu-yan;JIANG Chao;JIN Yan;ZHAO Yu-yang;State Key Laboratory of Dao-di Herbs Breeding Base,National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences;

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