呋喃唑酮代谢物间接竞争化学发光酶免疫法的建立  

Synthesis of furazolidone metabolite artificial antigen and development of indirect competitive chemiluminescence enzyme immunoassay

作　　者：黄登宇[1,2] 高文静[1] 黄种乾 高丽霞[1] 冯敏[1] 杨秀松[3] 

机构地区：[1]山西大学生命科学学院,山西太原030006 [2]山西大学食品药品快检技术中心,山西太原030006 [3]国家食品药品监督管理总局高级研修学院,北京100073

出　　处：《食品工业科技》2017年第9期294-299,共6页Science and Technology of Food Industry

摘　　要：建立快速检测动物源性食品中呋喃唑酮代谢物AOZ的间接竞争化学发光酶免疫法。利用对醛基苯甲酸(4-CBA)将AOZ衍生化为CPAOZ,利用活化酯法将CPAOZ与卵清蛋白(OVA)偶联,生成完全抗原CPAOZ-OVA。对包被原与抗体的最佳稀释倍数、封闭液浓度、竞争时间、酶标二抗稀释倍数进行优化,建立标准曲线,同时对方法的特异性进行评价。结果表明:包被抗原最佳稀释倍数为2000倍,抗体的最佳稀释倍数为20000倍、封闭液为2%脱脂乳粉、竞争时间为2 h、二抗最佳稀释倍数为4000倍;线性范围是0.075~7.396 ng/m L,半抑制浓度IC_(50)为0.74 ng/m L,对呋喃唑酮原药的交叉反应率为23.4%,与其他结构类似物及衍生化试剂的交叉反应率均小于0.1%。该方法灵敏度高、特异性好,可为监管部门对于快速检测动物源性食品中AOZ提供依据。An indirect competitive chemiluminescence enzyme immunoassay （ic-CLEIA）was used for determination and identification of the furazolidone metabolite in animal-derived food.AOZ can be translated into hapten CPAOZ by derivative reaction.CPAOZ-OVA,as the coating antigen, was prepared by activated ester method.After optimizing the dilution multiple of coating antigen and anti-body, blocking buffer, competitive reaction time and the dilution multiple of second antibody, a standard curve was established, and finally the specificity of the method was evaluated.The results indicated that, the optimum dilution multiple of coating antigen was 2000-fold,the anti-body was 20000-fold.Blocking buffer was dried skim milk at the concentration of 2%.The competitive reaction time was 2 h, and the ideal dilution multiple of secondary antibody was 4000- fold.The linearity range was 0.075-7.396 ng/mL, and half-inhibitory concentration （ICs0）value was 0.74 ng/mL.The crossreactivity value between AOZ and furazolidone original drug was 23.4% , and cross reactivity value with other analogues and derivatives was 〈 0.1%.The method with demonstrated excellent sensitivity and specificity can provide the basis for rapid detection of AOZ content in animal-derived food for supervision department.

关 键 词：呋喃唑酮 间接竞争化学发光酶免疫法 动物源性食品 快速检测 

分 类 号：TS207.3[轻工技术与工程—食品科学]