目的 构建一种能够可视化快速鉴定中药全蝎及其伪混品的方法。方法 应用改良十二烷基硫酸钠碱变性(SDS)法提取基因组DNA。选取细胞色素C氧化酶亚基Ⅰ(COⅠ)基因序列设计全蝎特异性引物,进行标记。优化聚合酶链式反应(polymerase chain reaction,PCR)体系,克隆制备全蝎阳性质控品作为对照。构建PCR产物检测试纸条,鉴定市售样品的真伪。结果 只有正品中药全蝎在286 bp处扩增出特异性条带,且试纸条的检测结果为阳性,检测灵敏度达到1.0×10-2 ng·μL-1。20个全蝎市售样品中18个正品样品,2个不合格样品。结论 建立的中药全蝎可视化试纸条检测的方法特异、灵敏、稳定、简单、快速,为中药全蝎的真伪鉴定提供了安全可行的方法。
Abstract
OBJECTIVE To establish a visual and rapid identification method for scorpion and its adulterants. METHODS Genomic DNA was extracted by improved SDS method. The COⅠ gene sequence was selected to design specific primers for scorpion and marked. Optimization of PCR reaction system, cloning preparation of scorpion positive quality control products as control. The PCR product test strip was constructed to identify the authenticity of commercial samples. RESULTS Only the scorpion of the genuine Chinese medicine amplified a specific band at 286 bp, and the detection results of the test strip were positive, with the detection sensitivity of 1.0×10-2 ng·μL-1. Among the 20 commercial scorpion samples, 18 were genuine and 2 were counterfeit. CONCLUSION The established method is specific, sensitive, stable, simple and rapid, which provides a safe and feasible method for the authenticity identification of Chinese medicine scorpion.
关键词
全蝎 /
改良十二烷基硫酸钠碱变性法 /
聚合酶链式反应 /
分子克隆 /
核酸试纸条
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Key words
scorpion /
fevised SDS method /
polymerase chain reaction /
molecular cloning /
nucleic acid test strip
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中图分类号：
R282
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脚注
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基金
吉林省科技发展计划项目资助(20200404152YY,20200403047SF);北华大学研究生创新计划项目资助(北华研创合字[2021]064)
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