甲基苯丙胺的胶体金免疫层析检测卡的研制

Development of a colloidal gold immuno-chromatography assay to detect methamphetamine

目的建立一种无需仪器、敏感、特异并适用于非专业人员现场快速检测甲基苯丙胺(冰毒)的方法。方法用冰毒-BSA免疫BALB/C小鼠,采用有限稀释法筛选特异性单克隆抗体并用分型试剂盒分析抗体亚型。采用物理方法将胶体金标记纯化的抗体,组装胶体金快速检测卡并检测此卡的灵敏度和特异性。结果采用有限稀释法筛选出7株特异性分泌株,其中R6腹水纯化后间接竞争ELISA测其效价为108,分型试剂盒测试显示此抗体属于IgG1型;金标抗体量为5μg/ml时,胶体金免疫检测卡的灵敏度为500ng/ml,与8种结构类似物均无交叉反应。结论高灵敏度的胶体金免疫层析快速检测卡作为毒品的初筛工具可有效地帮助查禁人员现场缉毒。

Objective A rapid detector method for non-professional staff detecting methamphetamine in scene,no instrument,sensitivity and specificity,would be set up.Methods Bagg albino C mice immunized with methamphetamine-modified bovine serum albumin were utilized for monoclonal antibody generation by the method of limited dilution technology.The detection of antibody typing kit was used for analysis the antibody subtype.Physical method was used for purified antibody labeled with colloidal gold.Colloidal gold test strip was composed and detected sensitivity and specificity.Results Seven special monoclonal antibodies were screened by limited dilution technology,among which valence of purified R6 antibody reached 108 via ciELISA,and belonged to IgG_1 by the detection of antibody typing kit.Monoclonal antibody against methamphetamine labeling with colloidal gold particles was 5μg/ml.The sensitivity of colloidal gold immuno-chromatography assay （GICA） for methamphetamine was 500ng/ml.The cross-reactivity of the GICA detection card to eight analogs did not appear.Conclusion GICA with high sensitivity,as a screening tool,may effectively contribute to Public Security Officers to detect narcotics in scene.