Jiang Chao, Yuan Yuan, Liu Libing, Hou Jingyi, Jin Yan, Huang Luqi
State Key Laboratory Breeding Base of Dao-di Herbs, National Resources Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, P.R. China.
Beijing Area Major Laboratory of Protection and Utilization of Traditional Chinese Medicine, College of Resources, Beijing Normal University, Beijing, 100875, P.R. China.
Sci Rep. 2015 Nov 5;5:16260. doi: 10.1038/srep16260.
A label-free, homogenous and sensitive one-step method for the molecular authentication of medicinal snakes has been developed by combining a rapid PCR technique with water-soluble cationic conjugated polyelectrolytes (CCPs). Three medicinal snake materials (Deinagkistrodon acutus, Zaocys dhumnades and Bungarus multicinctus; a total of 35 specimens) and 48 snake specimens with similar morphologies and textures were clearly distinguished by the naked eye by utilizing a CCP-based assay in a high-throughput manner. The identification of medicinal snakes in patented Chinese drugs was successfully performed using this detection system. In contrast to previous fluorescence-labeled oligonucleotide detection and direct DNA stain hybridization assays, this method does not require designing dye-labeled primers, and unfavorable dimer fluorescence is avoided in this homogenous method.
通过将快速聚合酶链反应（PCR）技术与水溶性阳离子共轭聚合物（CCP）相结合，开发出了一种用于药用蛇分子鉴定的无标记、均相且灵敏的一步法。利用基于CCP的检测方法，以高通量方式通过肉眼清晰区分了三种药用蛇材料（尖吻蝮、乌梢蛇和银环蛇；共35个样本）以及48个形态和质地相似的蛇样本。使用该检测系统成功对专利中药中的药用蛇进行了鉴定。与先前的荧光标记寡核苷酸检测和直接DNA染色杂交分析相比，该方法不需要设计染料标记引物，并且在这种均相方法中避免了不利的二聚体荧光。
