可视化微阵列蛋白芯片法同时检测蜂蜜中3种农药残留

目的建立可视化微阵列蛋白芯片法同时检测蜂蜜中多菌灵、啶虫脒、蝇毒磷3种农药残留含量的分析方法。方法依次向制备好的微孔板芯片里加入50μL标准品工作液和50μL相应的抗体,在25℃、600 r/min下反应30 min;再向每孔中加入50μL纳米银标记的羊抗鼠IgG, 37℃、600 r/min下反应30 min;最后显色并用芯片专用软件(MiELISA)进行自动化分析。结果该方法对多菌灵、啶虫脒、蝇毒磷的定量检测范围分别为0.4~6.4、0.3~4.8、0.75~12 ng/mL,相关系数r>0.99,检测限分别为3.21、2.47、2.52 ng/mL,准确度为83.6%~126.4%,变异系数均<10%,且三者之间具有极低的交叉反应;向不同种类的空白蜂蜜中添加2种不同浓度的标准品时,检测结果具有较高的准确度和重复性。结论本研究建立的可视化微阵列蛋白芯片检测法操作简便,检测限低,准确度、重复性、特异性均很好,且具有高通量、多指标同时检测等优点,适用于蜂蜜中多菌灵、啶虫脒、蝇毒磷3种农药残留同时检测。

Simultaneous determination of 3 kinds of pesticide residues in honey by visual microarray protein chip method

Objective To establish a method for the simultaneous determination of 3 kinds of pesticide residues in honey by visual microarray protein chip method.Methods The 50μL of standard working solution and 50μL of the corresponding antibody were added to the prepared microplate chip in sequence,and the reaction time was 30 min at 25℃and 600 r/min.Then 50μL of nano-silver-labeled goat antibody was added to each well Goat anti-mouse IgG,the reaction time was 30 min at 37℃and 600 r/min,the final color was developed and the chip-specific software(MiELISA)was used for automated analysis.Results The quantitative detection ranges of carbendazim,acetamiprid and coumaphos by this method were 0.4?6.4,0.3?4.8,0.75?12 ng/mL,respectively,and the correlation coefficient(r)was greater than 0.99.The detection limits were 3.21,2.47,2.52 ng/mL,respectively,the accuracy was 83.6%~126.4%,the coefficient of variation was less than 10%,and there was a very low cross reaction among them.When 2 different concentrations of standard substances were added to different types of blank honey,the detection results had high accuracy and repeatability.Conclusion The visual microarray protein chip detection method established in this study is easy to operate,has low detection limit,good accuracy,repeatability,and specificity,and has the advantages of high throughput and simultaneous detection of multiple indicators.It is suitable for simultaneous detection of 3 kinds of pesticide residues of carbendazim,acetamiprid and coumaphos in honey.