双抗夹心ELISA法检测双歧杆菌反应条件的优化  

Optimizing reaction conditions for Double-antibody Sandwich ELISA for Detection of Bifidobacterium spp

作　　者：庄翘楚[1] 孟祥晨[1] 

机构地区：[1]东北农业大学乳品科学教育部重点实验室食品学院,哈尔滨150030

出　　处：《中国乳品工业》2008年第5期55-58,共4页China Dairy Industry

基　　金：黑龙江省自然科学基金重点项目(ZJN03-3)

摘　　要：研究利用双抗夹心酶联免疫吸附法(Double-antibody Sandwich Enzyme-Linked Immunosorbent Assay,Double-antibody Sand-wich ELISA)快速检测双歧杆菌(Bifidobacterium)的最佳反应条件。使用实验室自制的兔抗两歧双歧杆菌免疫血清和鼠抗两歧双歧杆菌免疫血清,利用方阵法对两种血清抗体的反应浓度进行筛选,之后对包被液、封闭液、封闭时间及底物作用时间进行比较和选择,从而确定运用双夹心ELISA法快速检测双歧杆菌的最佳反应条件。最佳反应条件分别为:兔抗两歧双歧杆菌免疫血清和鼠抗两歧双歧杆菌免疫血清反应浓度分别为1︰1280,1︰320,包被液浓度0.05 mol/L(pH值为9.6)的Na2CO3-NaHCO3,封闭液为质量分数为5%的BSA-PBS,封闭30min,底物作用时间为30 min。优化了双抗夹心ELISA法检测双歧杆菌的反应条件,为双歧杆菌的血清学检测提供了参考。To optimize reaction conditions for Double-antibody Sandwich ELISA for Rapid Detection of Bifidobacterium were investigated. Anti-rabbit and anti-rat immune serums were made by our laboratory, The density of two immune serums in ELISA reaction was determined by using a square matrix, and then the coating buffer, confining hquid, confining time and substrate action time were optimized so as to determine the optimization reaction condition for Double-antibody Sandwich ELISA for Detection of Bifidobacterium. Optimization reaction conditions were as follows： the density of anti-rabbit and anti-rat immune serums were 1：1280 and 1：320 respectively; coating buffer was 0.05 mol/L pH9.6 Na2CO3-NaHCO3; confining liquid was 5%BSA-PBST, confining time was 30min; substrate action time was 30 rain. Reaction conditions for Double-antibody Sandwich ELISA for Detection of Bifidobacterium were optimized in this study, and this results provide a strategy to detect Bifidobacterium serologically.

关 键 词：双歧杆菌 双抗夹心ELISA 反应条件 

分 类 号：Q93-332[生物学—微生物学]