新霉素免疫膜层析检测方法研究

Development of an Immunochromatographic Strip Test for Detection of Neomycin

【目的】研制新霉素(neomycin,NEO)免疫膜层析快速检测试纸。【方法】建立分泌抗新霉素单克隆抗体(NEO mAb)杂交瘤细胞株,用胶体金免疫膜层析试验(GICA)制备NEO残留快速检测试纸(NEO-Strip),并对其性能进行鉴定。【结果】抗新霉素单克隆抗体的亲和力常数为3.75×1010 mol.L-1;制备的NEO-Strip目测限为50 ng.mL-1;试纸可特异性检测NEO,与庆大霉素、链霉素、妥布霉素、紫苏霉素、莱克多巴胺、沙丁胺醇、泰乐菌素和氯霉素等其它药物均无交叉反应性;就定性检测而言,其结果与高效液相色谱串联质谱法(HPLC-MS-MS)完全一致;不同的基质液对试纸的敏感度影响甚微。【结论】建立了特异的NEO免疫层析检测方法。

【Objective】 The objective of this experiment is to develop an immunochromatographic strip for rapid detection of neomycin（NEO）.【Method】 The monoclonal antibodies（mAb） against neomycin were generated and labeled with colloidal gold particles.Then the immunochromatographic strips for detection of NEO were developed and identified.【Result】 Ka of NEO monoclonal antibody was 3.75×1010 mol·L-1.The detection limit of the strip was 50 ng·mL-1 by naked eyes.The cross-reactivity with other antibiotics,such as gentamicin,streptomycin,tobramycin,sisomicin,ractopamine,salbutamal,tylosin,chloramphenic,was not detected.In parallel experiments,the test strip gave the same results as HPLC-MS-MS did.【Conclusion】 An immunochromatographic strip test method was established and the strip can be used to detect NEO residues rapidly with high specificity,sensitivity and simplicity in different samples.