间接竞争酶联免疫吸附法检测蜂王浆中链霉素

Competitive Indirect ELISA for Determination of Streptomycin in Royal Jelly

目的:基于定量检测蜂王浆中链霉素的残留量,建立快速检测蜂王浆中链霉素的间接竞争酶联免疫吸附方法。方法:采用制备链霉素人工抗原、筛选杂交瘤细胞、利用体内诱生腹水法制备抗链霉素单克隆抗体。结果:Logit/Log拟合标准曲线为y=-2.1x+0.8,相关系数r=0.9943,线性检测范围为4～324μg/kg,半数抑制浓度(IC50)为79μg/kg,检测限为4μg/kg,灵敏度为0.1μg/kg,蜂王浆样品的检测值符合率大于90%;采用方法二的样品前处理方法,链霉素阴性样品的添加回收率为67.7%～86.7%,样品重复检测结果的变异系数为4.3%～5.6%。结论:建立的间接竞争酶联免疫吸附法具有较好的特异性、检测符合率、回收率和重复稳定性,可用于蜂王浆中链霉素残留的检测。

A competitive ELISA kit for fast detection of streptomycin （SM） residues in royal Jelly was developed. SM complete antigens were prepared and monoclonal antibodies against streptomycin were produced by hybridoma technology and in vivo ascites induction method. Results showed that the kit＇s Logit/Log calibration curve was y = - 2.1x ＋0.8（correlation coefficient, r = 0.9943）, with linear range of 4 to 324 μg/kg. The half inhibitory concentration （IC50） was 79 μg/kg, sensitivity was 0.1 μg/kg, and limit of detection was 4 μg/kg. The detection accuracy was over 90%. The recoveries of SM from spiked positive samples were between 67.7% and 86.7% and the coefficients of variation for replicate measurements ranged from 4.3% to 5.6%. The developed ELISA kit can be applied for the quantitative detection of SM residues in royal jelly.