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目的研制间接竞争酶联免疫吸附法(ELISA)检测苯并芘试剂盒。方法采用棋盘滴定法确定抗原抗体的最佳用量;采用间接ELISA分析法确定苯并芘检测的最适条件,包括p H值、盐离子浓度及甲醇浓度。结果抗原最佳包被浓度为10μg/m L,单克隆抗体稀释倍数为1:5 000;在最优p H值为7.4、盐离子浓度为0.01 mol/L及甲醇浓度为10%的条件下,检测方法在5~50 ng/m L范围内线性相关,线性方程为y=-28.105 Ln(x)+123.66,R2=0.9937,IC50为13.74 ng/m L,苯并芘的最低检出限为3.3 ng/m L;在水样中的加标回收率为94.8%~112.3%;变异系数为4.38%~9.72%。结论该方法适用于水体中苯并芘的检测,可用于大批量样品的快速筛查。
Objective To develop an indirect competitive enzyme-linked immunosorbent assay (ELISA) for the detection of benzopyrene. Methods The optimum dosage of antigen and antibody was determined by chessboard titration. The optimum conditions of benzopyrene detection were determined by indirect ELISA, including p H value, salt ion concentration and methanol concentration. Results The optimum antigen concentration was 10 μg / ml and the dilution of monoclonal antibody was 1: 5 000. Under optimal pH value of 7.4, salt concentration of 0.01 mol / L and methanol concentration of 10% , The detection method was linear in the range of 5 ~ 50 ng / m L with the linear equation of y = -28.105 Ln (x) + 123.66, R2 = 0.9937 and IC50 of 13.74 ng / m L. The detection limit of benzopyrene Was 3.3 ng / m L. The spiked recoveries in water samples ranged from 94.8% to 112.3% and the coefficients of variation ranged from 4.38% to 9.72%. Conclusion This method is suitable for the detection of benzopyrene in water and can be used for rapid screening of large quantities of samples.