Zhou Yuxin, Nie Jing, Yu Shiqi, Hu Zhigang, Wang Bo
College of Pharmacy, Hubei University of Chinese Medicine, Wuhan, China.
Chinese Medicine Testing and Research Center, Hubei Institute for Drug Control, Wuhan, China.
Front Pharmacol. 2020 Apr 21;11:501. doi: 10.3389/fphar.2020.00501. eCollection 2020.
, an important traditional Chinese medicine, possesses remarkable medicinal activities, while lots of adulterants from other species were misused as for its large demand and resource starvation. In order to accurately identify and its common adulterants such as , , , and , a simultaneous identification method was designed with multiplex ligation-dependent probe amplification (MLPA) analysis. Five species-specific MLPA probe-couples for and its common adulterants were designed based on the universal primer amplified sequences, which can specifically detect the five species with no mutual interference, and sensitivity analysis showed as less as 5% or 8.75% adulterants in the mixed samples can be identified in a MLPA assay, especially, the relative quantity of the adulterants can be also inferred based on the MLPA peak area values. Moreover, the results of the present study confirmed the effectiveness of this technique in terms of simultaneous identification of and its common adulterants in an assay, which has great potential for ensuring the safety of this commercially valuable snake species.
某重要中药材具有显著药用活性，然而因其需求巨大且资源匮乏，大量来自其他物种的掺假品被误用作该药材。为准确鉴别该药材及其常见掺假品，如[具体掺假品名称1]、[具体掺假品名称2]、[具体掺假品名称3]和[具体掺假品名称4]，设计了一种基于多重连接依赖探针扩增（MLPA）分析的同时鉴别方法。基于通用引物扩增的[药材名称]序列设计了5对用于该药材及其常见掺假品的物种特异性MLPA探针，可特异性检测这5个物种且互不干扰，灵敏度分析表明，在MLPA检测中，混合样品中低至5%的[掺假品名称1]或8.75%的[掺假品名称2]等掺假品均可被鉴别，尤其还可根据MLPA峰面积值推断掺假品的相对含量。此外，本研究结果证实了该技术在一次检测中同时鉴别该药材及其常见掺假品的有效性，这对于确保这种具有商业价值的蛇类药材的安全性具有巨大潜力。
