食品中诺氟沙星残留管式化学发光免疫测定法的研究

Study on the tubular chemiluminescence immunoassay for the determination of norfloxacin residues in food

目的建立检测食品中诺氟沙星(norfloxacin,NFLX)残留的管式化学发光免疫测定法(chemiluminescence immunoassay,CLIA)。方法以磁性微粒为固相载体,通过优化抗原包被浓度、磁微粒稀释度、反应时间及pH值,建立一种可以检测食品中诺氟沙星残留的管式化学发光免疫测定法。结果该方法的检测范围为0.1668~68.6804 ng/mL,检测限为0.1161 ng/mL,准确度为94.52%,灵敏度为1.8717 ng/mL,变异系数<10%,NFLX与洛美沙星、氧氟沙星、恩诺沙星、环丙沙星交叉反应极低。结论本研究建立的NFLX-CLIA测定法特异性、灵敏度、准确度、重复性均符合要求,样本前处理简单,检测时间短,适用于食品中诺氟沙星残留的检测。

Objective To establish a method for the determination of norfloxacin(NFLX) by tubular chemiluminescent immunoassay(CLIA) with high sensitivity and specificity. Methods Magnetic particles were used as solid phase carrier to establish the CLIA method for the determination of norfloxacin residues in food by optimizing the concentration of antigen coating,dilution of magnetic particles,reaction time and pH value. Results The NFLX-CLIA showed an excellent performance with a liner detection range of 0.1668~68.6804 ng/mL,the detective limit of 0.1161 ng/mL,the accuracy of 94.52%,the sensitivity of 1.8717 ng/mL,and variation coefficient of less than 10%. The cross reaction of NFLX with lomefloxacin,ofloxacin,enrofloxacin and ciprofloxacin was very low. Conclusion The NFLX-CLIA assay established in this study has high specificity,sensitivity,accuracy and repeatability. The sample pretreatment is simple and the detection time is short. It is suitable for the determination of norfloxacin residues in food.