Detection Of Three Mycotoxins By Immunochromatography Method Based On Gold Nanosphere And Gold Nanoflower Labeling

In this paper,gold nanospheres?AuNSs?and gold nanoflowers?AuNFs?particles were prepared by sodium citrate reduction method and gold seed growth method,respectively.Based on the antigens and antibodies prepared in this laboratory,an immunochromatography method based on AuNS labeled antigen was established to detect vomiting toxin.The detection of zearalenone by immunochromatography method based on AuNF labeling was established.A highly sensitive quantitative detection of aflatoxin M₁ by immunochromatography method based on AuNF labeling was established.1.Detection of DON by immunochromatography based on AuNS labeled antigenDeoxynivalenol gold nanosphere immunochromatographic test card?DON-GICT?was prepared by labeling antigen with spherical nanogold.After systematic parameter optimization study,the sensitivity of DON-GICT detection of DON standard reached50ng/mL,the detection time was about 10min,and the detection limit of the actual sample was 1000?g/kg,which had better anti-matrix interference effect.34 food and feed samples were tested,7 samples showed positive,and the remaining samples showed negative.Compared with the commercial DON-ELISA test results,the total coincidence rate was 94.1%.2.Preparation of gold nanoflowers and immunochromatographic assay for detection of zearalenoneCharacterized by transmission electron microscopy and other means,optimized by orthogonal test method,1#gold nanoflowers with a particle size of?116.7±2.0?nm and many branches were prepared.The conditions of gold nanoflower-labeled zearalenone monoclonal antibody were optimized,and a zearalenone gold nanoflower immunochromatographic test card?ZEN-GFICT?was established for qualitative detection of zearalenone in food and feed.The results showed that the detection sensitivities of ZEN-GFICT?contrast method?and?cancellation method?for ZEN standards were 0.5 and 6ng/mL,respectively.It was 6 times?contrast method?and 4.5times?cancellation method?of the traditional ZEN nano gold immunochromatographic test card?ZEN-GICT?.The total conformity of the ZEN-GFICT judgment results?contrast method and cancellation method?of 21samples compared with those of the ZEN-ELISA kit were 90.5%and 90.5%,respectively.3.A highly sensitive quantitative detection of aflatoxin M₁?AFM₁?by immunochromatography method based on gold nanoflower labeledA gold nanoflower immunochromatography method was established to quantitatively detect aflatoxin M₁ in milk by nonlinear four-parameter fitting curve.The effects of pH,buffer type,ionic strength and surfactant on immunochromatographic reactions were optimized.The time range of AFM₁ gold nanoflower immunochromatographic test card?AFM₁-GFICT?reading signal was determined to be 20-25min.When the detection range of AFM₁ was 20-1000pg/mL,the correlation coefficient of nonlinear four-parameter fitting curve was R²=0.9993and the minimum detection limit was 12.3pg/mL.The recovery of blank milk samples with AFM₁ standard concentration of 0.1,0.3 and 0.5?g/kg was measured from 80.4%to 118.2%,and the coefficient of variation?n=10?was from 4.27%to 9.43%.15 milk samples were detected by AFM₁-GFICT,and the results were compared with the results of commercial AFM₁-ELISA.The correlation coefficient was R²=0.9768.