在建立磺胺甲噁唑单克隆抗体(SMZmAb)杂交瘤细胞株和阻断ELISA方法的基础上,研制出SMZ残留快速检测试剂盒(SMZ-Kit),并对其性能进行了测定.结果表明,SMZ-Kit的标准曲线呈典型的S型,符合参数logit曲线拟合,相关系数R2=0.990 1,检测范围为1.0～128.0 μg/L,灵敏度为0.73μg/L,半数抑制浓度(IC50)为11.5 μg/L,检测限为1.0μg/L;饲料样、牛奶样的平均添加回收率为83.9%,83.3%,平均批内和批间变异系数均<15%;SMZ-Kit与磺胺嘧啶的交叉反应(CR%)为2.88%,与其他磺胺类药物无交叉反应;试剂盒在℃可保存6个月.
刘宣兵
,
张改平
,
王斌
,
滕曼
,
邓瑞广
,
侯玉泽
,
邢广旭
,
杨继飞
. 磺胺甲噁唑残留检测阻断ELISA试剂盒的研制[J]. 华北农学报, 2009
, 24(2)
: 214
-217
.
DOI: 10.7668/hbnxb.2009.02.044
A ELISA kit for detection of Sulfamethoxazole (SMZ-Kit)was developed with generating hybridoma lines that secrete SMZ monoclonal antibodies (SMZmAb) and the method of blocking ELISA,its traits were identified. The calibration curve of SMZ-Kit was typical sigmoid curve fitted to the four parameter logistic equation with the linear detection of 1.0 to 128.0μg/ L (R2 = 0.9901),the sensitivity of 0.73μg/ L,the IC50 of 11.5μg/ L and the detection limit of 1.0μg/L.The recoveries of SMZ spiked in feed were 83.9 % and in milk were 83.3 %.The precision and accuracy of the assay as determined by inter-assay and intra-assay coefficient variation were below 15 %.The SMZ - Kit had 2.88 % cross-reactivity towards Sulfadiazine (SD) and little or no cross-reactivity towards other Sulfonamides.The validity of SMZ-Kit in 4℃was above six months.
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