蕲蛇DNA检测试剂盒的评价及应用

Evaluation of Agkistrodon acutus(Güenther) DNA Test Kit and Its Application in Quality Inspection of Commercial Products

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 kind of Agkistrodon acutus （Gtienther） DNA test kit, evaluate its quality indexes including specificity, stability, sensitivity and repeatability, and inspect the qualities of commercial Agkistrodon acutus （Giienther） samples. METHODS The Agkistrodon acutus （Güenther） DNA test kit was developed and modified according to the method recorded in Chinese Pharmacopoeia （2010 edition）. Eighteen Agkistrodon acutus （Gtienther） samples were randomly collected from Beijing, Tianjin, and Changchun. The kit assay was performed to identify these samples with the pharmacopoeia method as the standard control. RESULTS The kit proved effective after 20 times of freezing and thawing, and repeatability test showed same data for three tests. The lower limit of quantitation was 0. 025 g. The specificity test confirmed that 14 samples were genuine, and 4 were adulterants. All of the identification results by the kit assay were in accordance with the ones by the pharmacopoeia method. CONCLUSION The developed DNA test kit is accurate and effective for identification of Agkistrodon acutus （Gtienther）. Compared with the pharmacopoeia method, it is simpler and more rapid, demonstrating broad prospect in quality inspection of Agkistrodon acutus （Gtienther）.