CAJ | 학술논문

[目的]建立高灵敏度的呋喃它酮酶联免疫检测方法．[方法]以戊二醛法将呋喃它酮代谢物5-甲基吗啉-3-氨基-2-唑烷基酮（ AMOZ）与牛血清白蛋白（ BSA）和卵清白蛋白（ OVA）偶联，再由NaBH4还原西佛碱得到结构稳定的免疫原AMOZ-BSA和AMOZ-OVA；用对醛基苯甲酸（ CPA）对AMOZ进行衍生化，得到衍生物CPAMOZ，由质谱鉴定，经N-羟基琥珀酰亚胺活化酯法将CPAMOZ与BSA和OVA偶联得到免疫原CPAMOZ-BSA和包被原CPAMOZ-OVA．经紫外扫描鉴定表明人工抗原合成成功．免疫Balb/c小鼠，ic-ELISA方法测定鼠血清多克隆抗体对AMOZ和NPAMOZ的特异性．[结果和结论]结果显示CPAMOZ-BSA免疫制备的抗体对NPAMOZ有良好的特异性．抗体效价为1∶32×104，对NPAMOZ的半抑制质量浓度为4．53 ng· mL-1；抗血清与CPAMOZ、AMOZ及呋喃它酮的交叉反应率为78．6％、1．5％及4．6％；与其他3种硝基呋喃类原药及其代谢物对硝基苯甲醛、对醛基苯甲酸等结构相似物的交叉反应均小于0．01％．
[목적]건립고령민도적부남타동매련면역검측방법．[방법]이무이철법장부남타동대사물5-갑기마람-3-안기-2-서완기동（ AMOZ）여우혈청백단백（ BSA）화란청백단백（ OVA）우련，재유NaBH4환원서불감득도결구은정적면역원AMOZ-BSA화AMOZ-OVA；용대철기분갑산（ CPA）대AMOZ진행연생화，득도연생물CPAMOZ，유질보감정，경N-간기호박선아알활화지법장CPAMOZ여BSA화OVA우련득도면역원CPAMOZ-BSA화포피원CPAMOZ-OVA．경자외소묘감정표명인공항원합성성공．면역Balb/c소서，ic-ELISA방법측정서혈청다극륭항체대AMOZ화NPAMOZ적특이성．[결과화결론]결과현시CPAMOZ-BSA면역제비적항체대NPAMOZ유량호적특이성．항체효개위1∶32×104，대NPAMOZ적반억제질량농도위4．53 ng· mL-1；항혈청여CPAMOZ、AMOZ급부남타동적교차반응솔위78．6％、1．5％급4．6％；여기타3충초기부남류원약급기대사물대초기분갑철、대철기분갑산등결구상사물적교차반응균소우0．01％．
[Objective]To establish a high sensitivity enzyme-linked immunosorbent assay for the detection of furaltadone .[Method]Furaltadone metabolite ( 3-amino-5-morpholinomethyl-2-oxazolidinone , AMOZ ) was coupled to carrier proteins ( bovine serum albumin or ovalbumin ) and reduced with NaBH 4 for the pro-duction of immunogen AMOZ-BSA and coating antigen AMOZ-OVA through glutaraldehyde method .With 4-carboxybenzaldehyde for derivatization reagent , AMOZ was made into CPAMOZ , which was successful-ly conjugated to carrier proteins according to the active ester method to form CPAMOZ-BSA, CPAMOZ-OVA.UV scanning showed that antigens were successfully linked to carrier proteins .After immunizing animal ( Balb/c mice) , polyclonal antibody against NPAMOZ was produced .[Result and conclusion]Antibody was diluted at 1∶32 ×104 and the IC50 value was 4.53 ng· mL-1 .The cross-reactivity ( CR) of the antibody with CPAMOZ, AMOZ and furaltadone was 78.6%, 1.5%and 4.6%respectively.There is almost no CR with other three nitrofurans and their metabolites , which indicates a high selectivity of the antibody .