摘要
为快速检测水产品中FZ残留ELISA ;将呋喃唑酮 (Furazolidone ,FZ)与牛血清白蛋白 (BSA)、人血清白蛋白 (HSA)联接 ,分别作为免疫原及包被原 ,建立了水产养殖动物组织中呋喃唑酮的ELISA筛选方法。实验结果表明 ,理想的呋喃唑酮———人血清白蛋白 (FZ- HSA )包被抗原的浓度为 1.2 5 μg/mL ,酶标二抗 (羊抗兔IgG HRP)的工作浓度为 1∶10 0 0 ,多抗 (FZ- PcAb)的工作浓度为 1∶3 2 0 0 ,可测定的最适范围为 10～ 10 0ng/mL ,最小检测限为 1ng/mL。本实验所建立的ELISA方法的孔间差异为 4.16% ,板间差异为 9.2 0 % ,实验重复性好。添加回收率试验测的虾肉样本品均回收率为 (94.86± 9.5 5 ) %。在实验浓度范围内 ,呋喃唑酮 (FZ)的抗血清与氯霉素 (Chloramphenicol)、磺胺二甲基嘧啶 (Sulfamethazine)、红霉素 (Erythromycin)、恩诺沙星 (Enrofloxacin)、甲氧苄啶 (Trimethoprim )等可能会因同时使用而导致共同残留的抗微生物药物未显示免疫交叉反应性 ,与结构相似或相近药物呋喃西林 (Furacilinum)的交叉反应性为 43 %。这表明所建立的方法有较高特异性和灵敏度 ,适合呋喃唑酮残留的快速筛选。整个测定时间为 5h。
FZ hapten was coupled to BSA and HSA by an improved Diazotization procedure. The antibodies were obtained successfully by immunizing New Zealand rabbits with FZ-BSA. FZ-HSA was artificial coating antigen. The indirect competitive ELISA for FZ was developed. No cross-reactivity was seen between antibody and five other anitbiotics including chloramphenicol, sulfamethazine, erythromycin, enrof loxacin and trimethoprim. The antibody showed that the cross-reactivity for furacilinum was no more than forty-five percent. The results of the experiment demonstrated that the optimal concentrations of the coating antigen, polyclonal antibody against FZ and sheep anti-rabbit IgG were 1.25μg/mL, 1∶3 200 and 1∶1 000, respectively. A standard curve was obtained,the regression equation was y=95.381-20.203x (R2=0.991 3). The average recovery rate of shirmp muscle by ELISA was 94.86%±9.55%. So an indirect competitive enzyme-linked immunosorbent assay for FZ was developed. The whole assay took 5 hours to complete.
出处
《中国海洋大学学报（自然科学版）》
CAS
CSCD
北大核心
2005年第2期213-218,共6页
Periodical of Ocean University of China
基金
农业部"948"项目课题项目 (2 0 0 3 Q0 9 0 5)资助
