Rapid Immunological Detection Of Copper Ions Based On CLEIA And FICA

[Objective]Use the prepared anti copper McAb and the synthesized copper antigen to establish the indirect competitive chemiluminescence linked enzyme immunoassay?CLEIA?and fluorescence immunochromatographic assay?FICA?for simple,fast and quantitative detection of copper content in herbal plants and soil samples.[Methods]Use the anti copper McAb and copper antigen to establish the indirect competitive CLEIA and FICA.The detection conditions of the two methods were optimized,the accuracy,precision,specificity,stability and the add recovery rate were evaluated as well.A total of 31 samples include herbal plants and soils were collected and pre-treated by microwave digestion combined with precipitation.The copper content was detected by the two immunological methods and ICP-AES after sample preparation,and the relevance between immunological methods and instrumental method was evaluated.[Results]?1?The copper detection sensitivity of indirect competitive CLEIA was17.9 ng/ml,the low detection limit was 1.9 ng/ml,and the linear range was 3.0 to 200ng/ml.The recovery rate of accuracy test was 98.8%to 105.5%,and the RSD was less than or equal to 5%.Specific test showed that the anti copper McAb had a slight cross reactivity rate of 13.3%and 3.3%to Mg²⁺and Zn²⁺.Storage the CLEIA microplates at-20?away from light for 2 to 8 weeks,the stability test showed no significant change in6 weeks.?2?The copper detection sensitivity of FICA was 3.6 ng/ml,the low detection limit was 0.2 ng/ml,and the linear range was 0.4 to 50 ng/ml.The recovery rate of intra-assay accuracy test was 93.5%¹08.7%,and the RSD was less than 15%.Meanwhile,the recovery rate of inter-assay accuracy test was 92.5%¹12.5%,and the RSD was less than 10%.Specific test showed that the anti copper McAb had a slight cross reactivity rate of 14.2%and 5.3%to Mg²⁺and Zn²⁺.Accelerated aging test indicated that the test strips were valid for 2 months at dry room temperature.?3?Herbal plants and soil samples were pre-treated and detected by the two immunological methods and ICP-AES for copper content.The two types of samples detected by indirect competitive CLEIA and ICP-AES showed good relativity with the correlation coefficient value of 0.972 and 0.989.At the same time,the two types of samples detected by FICA and ICP-AES showed good relativity with the correlation coefficient value of 0.924 and 0.969 as well.The above results indicated that the two immunological methods can be applied to the detection of actual environmental samples.[Conclusion]Successfully established the indirect competitive CLEIA and FICA for simple,fast and quantitative detection of copper content in various environmental samples,and the detection sensitivity,detection limit and various parameters of the two immunological methods are in line with the requirements of national inspection technical.