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目的研制一种蕲蛇DNA检测试剂盒,对试剂盒的特异性、稳定性、敏感性和重复性进行评价,并对市售蕲蛇药材进行质量考察。方法以《中国药典》(2010年增补版)蕲蛇DNA检测方法为基础进行改良,研制标准DNA检测试剂盒,随机抽取北京、天津、长春和吉林4个城市18个蕲蛇样品,采用试剂盒和药典方法进行检测。结果蕲蛇DNA检测试剂盒经反复冻融20次后依然有效。试剂盒的最低检测样品量为0.025 g,18个蕲蛇样品中14个样品为正品,4个样品为不合格品。3次重复性检测均可完全重现相同结果。试剂盒与药典法检测结果一致,特异性为100%。结论蕲蛇DNA检测试剂盒对蕲蛇药材鉴别准确有效,并且与药典法相比更加简单、快捷,这种方法在蕲蛇药材质量考察方面具有广阔的应用前景。
Objective To develop a viper snake DNA test kit, to evaluate the specificity, stability, sensitivity and repeatability of the kit, and evaluate the quality of the viper snake. Methods Based on the “Chinese Pharmacopoeia” (2010 supplement) viper DNA detection method as a basis for improvement, the development of a standard DNA test kit, randomly selected from Beijing, Tianjin, Changchun and Jilin 4 cities 18 viper samples, using a kit And pharmacopoeia testing. Results Viper DNA detection kit after repeated freezing and thawing 20 times still valid. The minimum test sample volume was 0.025 g, 14 of 18 samples were authentic and 4 samples were unqualified. The same result can be completely reproduced by 3 repetitive tests. Kit and pharmacopoeia test results, the specificity of 100%. Conclusion Viper DNA detection kit is accurate and effective for the identification of Agkistrodon acutus, and is simpler and faster than the pharmacopoeia. This method has broad application prospect in the quality inspection of Agkistrodon.