文档介绍：中国农业科学 2011,44(11):2387-2397
Scientia Agricultura Sinica doi: .0578-
新霉素免疫膜层析检测方法研究
王爱萍 1, 李发弟 1, 胡骁飞 2, 史兆国 1, 周景明 2, 杜晓明 2, 王林林 2, 禹乐乐 2, 刘明阳 2
(1 甘肃农业大学动物科学学院,兰州 730070;2 河南省农业科学院免疫学重点实验室, 郑州 450002)
摘要:【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 ×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***、沙丁***醇、泰乐菌素
和***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。
关键词:新霉素;单克隆抗体;胶体金;试纸条
Development of an Immunochromatographic Strip
Test for Detection of Neomycin
WANG Ai-ping1, LI Fa-di1, HU Xiao-fei2, SHI Zhao-guo1, ZHOU Jing-ming2, DU Xiao-ming2,
WANG Lin-lin2, YU Le-le2, LIU Ming-yang2
(1College of Animal Science, Gansu Agricultural University, Lanzhou 730070; 2Henan Key Lab for Animal Immunology,
Zhengzhou, 450002)
Abstract: 【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 ×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.
Key words: neomycin; monoclonal antibody; colloidal gold particles; test st