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磺胺二甲基嘧啶酶联免疫快速检测试剂盒
(SM2-ELISA)
1 概要
磺胺二甲基嘧啶（SM2）是应用广泛的抗生素，对于畜禽和动物水产品养殖过程中疾病的控制和治疗起到重要作用。研究证明SM2具有潜在的毒副作用，在动物机体中长期存在的SM2会发生多种代谢作用：代谢物主要为N4位置上的酰化化合物，N4位置上的葡萄糖化合物，和脱芳氨基化合物。若与亚硝酸盐共同摄取能够促进生成脱芳氨基代谢物的形成，该物质有一定的诱导突变作用。我国农业部第235号文件规定了其残留量为在牛奶中为25μg/kg，由于磺胺二甲基嘧啶的广泛应用，对于其他动物组织检测也有必要。由于高效液相色谱等仪器分析方法样本前处理盒测定的繁琐和费用高，推广使用受到了限制。
本试剂盒可用于定量、定性检测鱼、虾样本，动物组织，禽蛋中和牛奶样本中的SM2残留。与仪器分析技术相比较具有快速，简便，准确和灵敏度高，特异性好等优点。操作时间仅需1小时左右，同时能够实现多个样本的同时测定。在提供准确结果的同时，减少了工作强度。
2 检测原理
本试剂盒采用间接竞争抑制法检测样本中的SM2。在微孔板上预包被SM2抗原，样本或标准品中的SM2残留和包被抗原竞争结合抗SM2的单克隆抗体，经过孵育后，洗涤未与包被抗原结合的抗SM2的单克隆抗体，加入抗鼠单抗的二抗-HRP，经过孵育后，洗涤游离的二抗-HRP后用H2O2-TMB底物系统显色，样本吸光度值与其所含SM2的成负相关，与标准品溶液的比较即可得出SM2含量。
3 试剂盒组份
每个试剂盒中的试剂可进行48个测量（包括标准测定），试剂盒组分：
包被有SM2抗原的微孔条······················································· 48孔（8孔/条×6条）
板架················································································· 1块（8×12）
SM2标准品溶液·················································································· 4瓶（0.3ml/瓶）
分别为0号：0ppb；1号：0.4ppb；2号：2ppb；3号：10ppb。
抗SM2单克隆抗体浓缩液（100×）················································· 1瓶（50μL/瓶）
二抗-HRP浓缩液（100×）······························································· 1瓶（50μL/瓶）
抗体稀释液························································································· 1瓶（10ml/瓶）
底物溶液A··························································································· 1瓶（4 mL/瓶）
底物溶液B···························································································· 1瓶（4 mL/瓶）
终止液（含2 M的硫酸）··································································· 1瓶（4 mL/瓶）
20×浓缩洗涤液··················································································· 1瓶（25 mL/瓶）
10×样品复溶液··················································································· 1瓶（10 mL/瓶）
封板膜··································································································· 3张
说明书··································································································· 1份
4 另外所需试剂和仪器
4.1 试剂
--甲醇 （分析纯）
--正己烷（分析纯）
4.2 设备
--微孔板酶标仪（450 nm）
--恒温箱 （37℃）
--天平（精度0.01 g）
--匀浆器
--振荡器
--超声波清洗器（工作频率不小于40KHZ）
--离心机和离心管
--旋转蒸发器或氮气吹干仪
--可调式0.5μL-20 μL， 20 μL-200 μL 和100 μL-1000 μL 微量移液器
--多道移液器
4.3 耗材
10 mL圆底带盖离心管