[ Objective] Vibrio parahaemolyticus was chosen as the material to make rabbit anti- V. parahaemolyticus polyclonal antibody, the immuno-gold chromatography assay （IGCA） was used to develop immunogold labeling test ...[ Objective] Vibrio parahaemolyticus was chosen as the material to make rabbit anti- V. parahaemolyticus polyclonal antibody, the immuno-gold chromatography assay （IGCA） was used to develop immunogold labeling test strips for detecting V. parahaemolyticus. [Methods] Rabbit anti-V, parahaemolyticus IgG-2 was used to wrap nitrocellulose membrane detection line, goat anti-rabbit IgG was used to wrap control line, and im- muno-gold was used to label rabbit anti-V, parahaemolyticus IgG-1 to establish the immuno-gold chromatography assay （IGCA） for detection of V. parahaemolyticus. [Results] Test strip detection lines of positive results along with the control line were all red, test results could be got in only five to fifteen minutes, the minimum detectable amount of this method to V. parahaemolyticus was 3.60 x 104 cfu/ml, and cross reactions wouldnt occur when detecting common intestinal bacteria like Vibrio alginolyticus, vibrio cholerae, Vibrio damsela, Vibrio metschnikovii, Citrobacter freundii and salmonella etc. The test results were undifferentiated when the test strips were stored at 4 ~C for six months, stored at room temperature for three months or stored at 37 ~C for one month. [ Condusionl The established test strip assay was simple and rapid during operation, with high sensitivity, strong specificity and good stability, the test results were easy to be observed and judged, and this assay was very suitable for grassroots breeding department application.展开更多
从沙田柚自交花粉管中提取与自交不亲和相关蛋白,应用免疫胶体金标记抗体技术检测沙田柚Citrus grandis var. Shatinyu Hort.自花授粉后3 d 1/2花柱中花粉管特异蛋白的定位分布。结果表明,花粉管特异蛋白在花粉管中依次定位于内质网、...从沙田柚自交花粉管中提取与自交不亲和相关蛋白,应用免疫胶体金标记抗体技术检测沙田柚Citrus grandis var. Shatinyu Hort.自花授粉后3 d 1/2花柱中花粉管特异蛋白的定位分布。结果表明,花粉管特异蛋白在花粉管中依次定位于内质网、细胞质和细胞纤维壁上。展开更多
稻曲病菌在PD 液体培养基中生长良好,并能产生对植物细胞具有高度生物抑制活性的毒素。生物学活性测定袁明,用100%的甲醇能提取稻曲病菌液体培养物中的粗毒素。粗毒素对小麦胚根胚芽的生长有强烈的抑制作用。把毒素主要成分Ustiloxin A ...稻曲病菌在PD 液体培养基中生长良好,并能产生对植物细胞具有高度生物抑制活性的毒素。生物学活性测定袁明,用100%的甲醇能提取稻曲病菌液体培养物中的粗毒素。粗毒素对小麦胚根胚芽的生长有强烈的抑制作用。把毒素主要成分Ustiloxin A 和BSA 偶联后,制备了抗血清,ELISA 检测表明用两种偶联剂偶联所制备的抗体效价分别为1∶20000和1∶6000。进一步的免疫胶体金标记分析表明,所制备的抗体能与茼丝中分泌的毒素特异性结合,说明所获得的抗体是特异性的。展开更多
A novel, sensitive anodic stripping volammetric immunoassay was developed based on silver-enhanced immuno-gold label. The immunoreaction was performed in a polystyrene microwell by using the sandwich format. Primary a...A novel, sensitive anodic stripping volammetric immunoassay was developed based on silver-enhanced immuno-gold label. The immunoreaction was performed in a polystyrene microwell by using the sandwich format. Primary antibodies specific for C-3(complement Ⅲ) were adsorbed passively on the walls of a polystyrene microwell. The C-3 analyte was first captured by the primary antibody and then sandwiched by a secondary colloidal gold-labeled antibody. The addition of the silver enhancement solution results in the precipitation of a large amount of silver on colloidal gold labels due to the catalytic reduction which, after silver metal dissolution in an acidic solution, was determined by anodic stripping voltammetry(ASV) at a glassy-carbon electrode. The influence of some immunoassay conditions upon the anodic stripping peak current was examined and optimized. The anodic stripping peak current depended linearly on the logarithm of C-3 mass concentration over the range of 7.2 ng/mL to 7.33 μg/mL and a detection limit as low as 7ng/mL is achieved. The anodic stripping volammetric immunoassay was applied to the determination of C-3 concentration in human serum with satisfactory results.展开更多
基金funded by the Science and Technology Projects of the State Administration of Quality Supervision,Inspection and Quarantine(2010IK016)
文摘[ Objective] Vibrio parahaemolyticus was chosen as the material to make rabbit anti- V. parahaemolyticus polyclonal antibody, the immuno-gold chromatography assay （IGCA） was used to develop immunogold labeling test strips for detecting V. parahaemolyticus. [Methods] Rabbit anti-V, parahaemolyticus IgG-2 was used to wrap nitrocellulose membrane detection line, goat anti-rabbit IgG was used to wrap control line, and im- muno-gold was used to label rabbit anti-V, parahaemolyticus IgG-1 to establish the immuno-gold chromatography assay （IGCA） for detection of V. parahaemolyticus. [Results] Test strip detection lines of positive results along with the control line were all red, test results could be got in only five to fifteen minutes, the minimum detectable amount of this method to V. parahaemolyticus was 3.60 x 104 cfu/ml, and cross reactions wouldnt occur when detecting common intestinal bacteria like Vibrio alginolyticus, vibrio cholerae, Vibrio damsela, Vibrio metschnikovii, Citrobacter freundii and salmonella etc. The test results were undifferentiated when the test strips were stored at 4 ~C for six months, stored at room temperature for three months or stored at 37 ~C for one month. [ Condusionl The established test strip assay was simple and rapid during operation, with high sensitivity, strong specificity and good stability, the test results were easy to be observed and judged, and this assay was very suitable for grassroots breeding department application.
文摘A novel, sensitive anodic stripping volammetric immunoassay was developed based on silver-enhanced immuno-gold label. The immunoreaction was performed in a polystyrene microwell by using the sandwich format. Primary antibodies specific for C-3(complement Ⅲ) were adsorbed passively on the walls of a polystyrene microwell. The C-3 analyte was first captured by the primary antibody and then sandwiched by a secondary colloidal gold-labeled antibody. The addition of the silver enhancement solution results in the precipitation of a large amount of silver on colloidal gold labels due to the catalytic reduction which, after silver metal dissolution in an acidic solution, was determined by anodic stripping voltammetry(ASV) at a glassy-carbon electrode. The influence of some immunoassay conditions upon the anodic stripping peak current was examined and optimized. The anodic stripping peak current depended linearly on the logarithm of C-3 mass concentration over the range of 7.2 ng/mL to 7.33 μg/mL and a detection limit as low as 7ng/mL is achieved. The anodic stripping volammetric immunoassay was applied to the determination of C-3 concentration in human serum with satisfactory results.
