可视化凝胶酶联免疫吸附分析法检测牛奶中庆大霉素和卡那霉素

建立了牛奶中同步测定庆大霉素和卡那霉素的可视化凝胶ELISA方法。在凝胶检测柱的两个检测层中填充CN-Br活化的Sephrose 4B凝胶-羊抗鼠Ig G作为固相载体,加入药物的单克隆抗体与载体结合,酶标抗原和待测样本中的药物共同竞争有限的单克隆抗体,通过3,3＇,5,5＇-四甲基联苯胺（TMB）底物显色定性判断药物的存在。本方法采用一步法,检测时间仅为15 min,对庆大霉素和卡那霉素的灵敏度为2.0μg/L,对牛奶中的两种药物检出限（Cut-off值）均为5μg/L。牛奶盲样比对实验表明,本方法与超高效液相色谱-串联质谱（UPLC-MS/MS）法测定结果相符。...

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Published in分析化学 Vol. 43; no. 6; pp. 881 - 885
Main Author 徐飞 栗静雅 周洁 刘茂林 刘义明 王建芬 丁双阳 李秀波
Format Journal Article
LanguageChinese
Published 中国农业科学院饲料研究所,国家饲料药物基准实验室,北京100081%中国农业大学动物医学院,国家兽药残留基准实验室,北京100094%北京市延庆县畜牧技术推广站,北京,102100 2015
Subjects
凝胶酶联免疫分析
卡那霉素
同步测定
庆大霉素
牛奶
庆大霉素
卡那霉素
Simultaneous detection
凝胶酶联免疫分析
牛奶
Gentamicin
Kanamycin
Gel-enzyme-linked immunosorbent assay
同步测定
Milk
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ISSN0253-3820
DOI10.11895/j.issn.0253-3820.141062

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Summary:建立了牛奶中同步测定庆大霉素和卡那霉素的可视化凝胶ELISA方法。在凝胶检测柱的两个检测层中填充CN-Br活化的Sephrose 4B凝胶-羊抗鼠Ig G作为固相载体,加入药物的单克隆抗体与载体结合,酶标抗原和待测样本中的药物共同竞争有限的单克隆抗体,通过3,3＇,5,5＇-四甲基联苯胺（TMB）底物显色定性判断药物的存在。本方法采用一步法,检测时间仅为15 min,对庆大霉素和卡那霉素的灵敏度为2.0μg/L,对牛奶中的两种药物检出限（Cut-off值）均为5μg/L。牛奶盲样比对实验表明,本方法与超高效液相色谱-串联质谱（UPLC-MS/MS）法测定结果相符。
Bibliography:Gel-enzyme-linked immunosorbent assay ; Gentamicin ; Kanamycin ; Simultaneous detection ;Milk
A visual gel-enzyme-linked immunosorbent assay （Gel-ELISA） assay for simultaneous detection of gentamicin （GEN） and kanamyein （KANA） in milk was described. In the gel detection column, two detection layers were filled with CN-Br activated Sephrose 4B gel as solid phase carrier, which was coupled with goat anti-mouse IgG. The anti-GEN and anti-KANA monoclonal antibodies were added to combine with the carrier respectively in each layer. Gentamicin and kanamycin drugs in sample competed with corresponding enzyme-labeled antigens for the immobilized antibody on the column, and the drugs could be detected by the chromogenic reaction of substrate 3,3＇,5,5＇-tetramethylbenzidine. This approach was a yes/no method after color detection by naked eyes with the detection time of 15 min. The sensitivity of the Gel-ELISA method was 2 μg/L, and the detection limit （ Cut-off value） for gentamicin and kanamycin in raw milk was 5 μg/L.
ISSN:0253-3820
DOI:10.11895/j.issn.0253-3820.141062