金黄色葡萄球菌B型肠毒素的酶联免疫检测

Enzyme Linked Immunosorbent Assay for Staphylococcal Enterotoxin B in Milk

[目的]为细菌毒素的检测提供理论依据。[方法]以BALB/c小白鼠为免疫对象,以自制的高纯度金黄色葡萄球菌B型肠毒素（SEB）为免疫抗原,研究其抗体的产生过程和效价,并建立SEB的间接竞争ELISA检测方法。[结果]自制SEB纯度较高,能较好地刺激动物产生抗体。以自制SEB为免疫抗原注射BALB/c小白鼠,其最高效价为1∶100 000。SEB浓度在1-1 000 ng/ml范围内,竞争抑制率与SEB浓度的对数值呈显著线性关系。当SEB的浓度为5-500 ng/ml时,随样品中SEB浓度的增加,污染样品的回收率变异系数减小,检测准确度提高。[结论]对不同污染程度的牛奶样品,利用间接竞争ELISA法检测其SEB残留的灵敏度远远高于生化法。

[Objective] The research aimed to provide theoretical basis for the detection of bacterial toxins.[Method] With BALB/C white mice as immune objects,the self-made staphylococcal enterotoxin B（SEB） with high purity was taken as immune antigen to study the producing process and titers of its antibodies and establish the indirect competitive ELISA detection method for SEB. [Result] The self-made SEB was of higher purity and it could stimulate animals to produce antibody.BALB/c white mice were injected with the self-made SEB and its highest titer was 1∶100 000.When SEB concentration was in the range of 1～1 000 ng/ml,the competitive inhibition rate showed a significant linear correlation with the logarithm value of SEB concn.When the concn.of SEB was 5～500 ng/ml,the recovery variation coefficient of the polluted samples was decreased and the detection accuracy was increased with the increase of SEB concn.in samples.[Conclusion] For milk samples with different pollution degree,the sensitivity of detecting SEB residues by using the indirect competitive ELISA method was greatly higher than that of biochemical method.