摘要
目的 建立基于特异性PCR技术的朝鲜淫羊藿及其同属近缘物种鉴别方法，为朝鲜淫羊藿药材鉴定与质量控制提供可靠的分子手段。方法 基于朝鲜淫羊藿基因组的微卫星（STRs）序列，设计特异性引物；对PCR反应体系中的聚合酶种类、退火温度、循环数、引物用量及不同PCR仪进行考察优化；验证方法的检出限、适用性。结果 针对朝鲜淫羊藿的STRs序列，设计的特异性引物可稳定扩增出214 bp的目标条带，对同属其他物种无扩增，检出限可达3.5 ng/μL。结论 建立的位点特异性PCR鉴别方法可快速、准确鉴别朝鲜淫羊藿及其同属近缘物种，为朝鲜淫羊藿的药材鉴定及质量控制提供了有效技术支撑。
关键词： 特异性PCR；朝鲜淫羊藿；同属物种；分子鉴别
Abstract
Objective This study aimed to develop a specific PCR-based method for the discrimination of Epimedium koreanum from its congeneric allied species, offering a robust molecular tool for the authentication and quality assurance of Epimedium koreanum herbal material. Methods Species-specific primers were designed targeting microsatellite (STRs) sequences in the Epimedium koreanum genome. A comprehensive optimization of the PCR reaction system was conducted, including the evaluation of polymerase types, annealing temperatures, cycle numbers, primer concentrations, and PCR instruments. The analytical performance of the method, including detection limit and applicability, was rigorously validated. Results The primers specifically designed for the STRs sequences of Epimedium koreanum yielded a consistent 214 bp amplification product, with no cross-reactivity observed in other congeneric species. The method demonstrated a low detection limit of 3.5 ng/μL. Conclusion The developed locus-specific PCR method enables rapid and accurate discrimination of Epimedium koreanum from its congeneric allied species, providing a reliable technical platform for the authentication and quality control of Epimedium koreanum herbal medicine.
Key words： Specific PCR; Epimedium koreanum; Congeneric species; Molecular identification
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