中成药蛤蚧定喘丸中原料动物药材的分子鉴定  

Molecular Identification of the raw animal medicinal materials of Chinese patent medicine Gejie Dingchuan Wan

作　　者：景晓彤 邓麒蓉 李宏宇 王乙淋 黄勇[1] JING Xiao-tong;DENG Qi-rong;LI Hong-yu;WANG Yi-lin;HUANG Yong(Guangxi University of Chinese Medicine,Nanning 530200,China;Guangzhou Huali Science and Technology Vocational College,Guangzhou,527324,China)

机构地区：[1]广西中医药大学,广西南宁530200 [2]广州华立科技职业学院,广东广州527324

出　　处：《时珍国医国药》2023年第7期1657-1660,共4页Lishizhen Medicine and Materia Medica Research

基　　金：广西自然科学基金(2020GXNSFAA238017,2023GXNSFDA026065,2017GXNSFDA198001);广西中医药大学科研项目(2018BS020);广西壮瑶药重点实验室项目(GXZYKF2022-20)。

摘　　要：目的建立蛤蚧定喘丸中原料动物药材的分子鉴定方法。方法首先构建了蛤蚧定喘丸中原料动物药材的16S rRNA序列的数据库;然后收集3个不同厂家(每个厂家3个不同批号)和自制的蛤蚧定喘丸共计12份,使用改良CTAB法对蛤蚧定喘丸进行总DNA提取,扩增16S rRNA,对PCR产物进行克隆测序,然后对测序结果进行BlastN分析。结果共挑取131个阳性克隆进行测序,序列去除载体序列和人工校正后,12份蛤蚧定喘丸样品均能鉴定出蛤蚧和鳖甲成分,其中鉴定结果为蛤蚧的有87个样品,占总样品的66.4%;鉴定结果为中华鳖的有44个样品,占总样品的33.6%。结论本研究的分子鉴别方法可鉴定蛤蚧定喘丸中原料动物药材,为该中成药的质量控制提供参考。Objective To establish a molecular identification method for the raw animal medicinal materials of Chinese patent medicine Gejie Dingchuan Wan.Methods Firstly,the 16S rRNA sequence database of raw animal medicinal materials of Gejie Dingchuan Wan was established;Then a total of 12 commercial Gejie Dingchuan Wans from three different manufacturers(each manufacturer has three different batch numbers)and self-made Gejie Dingchuan Wan.The total DNA of Gejie Dingchuan Wan was extracted by improved CTAB method,16S rRNA was amplified,the PCR products were cloned and sequenced,and the sequencing results were analyzed by BlastN.Results A total of 131 positive clones were selected for sequencing.After removing the vector sequence and manually correcting the sequence,12 Gejie Dingchuan Wans samples could identify the components of Gekko gecko and Trionyx sinensis,among which 87 Gekko gecko,accounting for 66.4%of the total samples and 44 Trionyx sinensis,accounting for 33.6%of the total samples.Conclusion The molecular identification method in this study can be used to identify the raw animal medicinal materials of Gejie Dingchuan Wan and provide reference for the quality control of this Chinese patent medicine.

关 键 词：蛤蚧定喘丸 蛤蚧 16S rRNA 分子鉴定 中成药 

分 类 号：R282.5[医药卫生—中药学]