百菌清残留阻断ELISA试剂盒的研制及鉴定

Development and authentification of a rapid competitive ELISA kit for Chlorothalonil residues

中文关键词:  百菌清  单克隆抗体  阻断ELISA  快速检测试剂盒

英文关键词:Chlorothalonil  monoclonal antibody  competitive ELISA  rapid test kit

中文摘要:

      目的 研制百菌清(Chlorothalonil, CTN)残留快速检测阻断ELISA试剂盒(CTN-Kit), 并对其特性进行测定。方法 基于一株分泌抗CTN高亲和力的CTN单克隆抗体(monoclonal antibody, mAb), 应用阻断ELISA试验原理研制CTN-Kit。结果 CTN-Kit的标准曲线呈典型的S型, 相关系数R2=0.9922, 符合4参数logit曲线拟合, 线性检测范围为1.03~845.46 μg/L, 灵敏度为2.5 μg/L, 半数抑制浓度(IC50)为29.44 μg/L, 检测限为3.0 μg/L; 黄瓜样和番茄样的平均添加回收率分别为92.0%、94.0%, 平均批内和批间变异系数均低于10%; CTN-Kit与五氯硝基苯的交叉反应率(CR%)为1.7%, 与其他竞争物几乎没有反应性; 试剂盒在4 ℃可保存6个月以上。结论 CTN-Kit灵敏度高、准确性、重现性好, 可用于农产品中CTN的残留检测。

英文摘要:

      Objective To develop a competitive ELISA test kit for the detection of Chlorothalonil (CTN) and determine its characteristics. Methods A competitive ELISA test kit for the detection of CTN (CTN-Kit) was developed based on a specific CTN monoclonal antibody (mAb). Results The calibration curve of the CTN-Kit was a typical sigmoidal curve, fitting a four parameter logistic equation over the range of 1.03 to 845.46 μg/L of CTN (R2=0.9922), with the sensitivity of 2.5 μg/L, IC50 of 29.44 μg/L and a detection limit of 3.0 g/L. The recoveries for CTN doping of cucumber and tomato were 92.0% and 94.0% respectively. The precision and accuracy of the assay as determined by inter-assay and intra-assay coefficient variation were both below 10%. The CTN-Kit gave a cross-reactivity of 1.7% with quintozene and little or no cross-reactivity towards other compounds. The viability of the CTN-Kit at 4 ℃ was above six months. Conclusion This CTN-Kit has a good sensitivity, accuracy and reproducibility, and is suitable for the detection of CTN in agricultural products.