畜牧兽医学报2011，Vol.42Issue(6)：857-864,8.
新霉素阻断ELISA试剂盒的研制与应用
Development and Evaluation of Blocking ELISA Kit for Neomycin Detection
摘要
Abstract
A blocking ELISA kit for detection of neomycin was developed based on the NEO mAb, and its properties were examined. The result showed that the calibration curve of NEO-kit was a typical sigmoid curve fitted to the four parameters logistic equation with the linear detection of 1.0 to 64.0 μg · L-1 and R2 was 0. 990 3. The kit has a sensitivity of 0. 75 μg · L-1 , the IC50 of 2. 59 μg · L-1and the detection limit of 1. 0 μg · L-1. The kit had no cross-reactivity with gentamicin, streptomycin, oxytetracycline, ciprofloxacin, difloxacin, salbutamol, sulfadiazine and sulfamethoxazole. The recoveries of NEO in milk, feed and meat were 89. 50％, 89.58％ and 87. 92％, respectively. The inter-assay and intra-assay coefficient variation were all below 15％.Moreover, the inter-assay coefficient variation was lower than that of intra-assay. Different sample diluted solution had very little effect on the NEO-kit. The kit remained effective for storing at 4 ℃ more than six month. There was no significant difference of the recovery of neomycin in milk between with NEO-kit and with HPLC-MS-MS (P≥0. 05). The neomycin content in feed determined by NEO-kit and HPLC-MS-MS has no significant difference ( P≥0. 05). In conclusion, we developed a NEO-kit with high sensitivity, specificity and accuracy.关键词
新霉素/单克隆抗体/阻断ELISA/试剂盒Key words
neomycin/ monoclonal antibody/ blocking ELISA: kit分类
轻工纺织引用本文复制引用
王爱萍,李发弟,胡骁飞,曾巧英,李金录,郭艳丽,赵启法,陈玉梅,范璐..新霉素阻断ELISA试剂盒的研制与应用[J].畜牧兽医学报,2011,42(6):857-864,8.基金项目
"十一五"国家科技支撑计划重大项目(2006BAK02A21) （2006BAK02A21）
