Study Of The Mycotoxin Detection Based On Homogeneous Immunoassay Technology

Mycotoxins are secondary metabolites produced by fungi,which are likely to emerge in the growth,store,transportation and processing of agricultural products.Most of the mycotoxins have carcinogenic,teratogenic and mutagenic toxicities,which seriously endanger the health of humans and cause great harm to the socio-economic.Therefore,the development of a highly sensitive,convenient and rapid method of detection of mycotoxins has very important research significance and application value.At present,the detection methods of mycotoxins mainly include thin layer chromatography?TLC?,high performance liquid chromatography?HPLC?,gas chromatography?GC?,mass spectrometry?MS?,enzyme-linked immunosorbent assay?ELISA?.The drawbacks of the TLC are that the sample preparation process is cumbersome;the disadvantages of the HPLC method are that the reagents and instruments required are expensive;the GC method has the disadvantages of high cost of testing equipment;the MS method often needs to be combined with HPLC or GC method,and the detection process is complex;the ELISA method is complex,and the quantitative analysis accuracy of this method is very poor,these shortcomings also restrict its application in the rapid detection of mycotoxins..The luminescent oxygen channeling immunoassay technology not only has the advantages of high sensitivity and accurate quantification,but also realizes one-step,wash-free detection,and the operation process is simple,of good stability,easy to achieve miniaturization and automated detection.In the manuscript,in order to achieve the needs of rapid detection of mycotoxins,we present a method based on luminescent oxygen channeling immunoassay technology for mycotoxins detection.First of all,we have figured out the principle of the method,and a homemade high-throughput homogeneous immunoassay system is constructed according to its measurement principle.Secondly,we studied the detection method about two common toxins,which are aflatoxin B₁and deoxynivalenol.The experimental method used is the competitive method,through the competitive effects of binding to monoclonal antibodies between the samples and biotinylated complete antigen to achieve quantitative detection.The third,according to the immunoreaction and homogeneous detection steps,we studied the optimal detection conditions of aflatoxin B₁ and deoxynivalenol respectively.Finally,a competitive method for the detection of mycotoxins based on luminescent oxygen channeling immunoassay technology was established.Experiments were carried out under optimized experimental parameters.The experimental results show that,for aflatoxin B₁,the method can achieve the lowest detection limit of 0.048ng/mL,the lowest limit of quantification of 0.22ng/mL,the intra-and inter-assay variation both below 10%,and the recovery rate is within 87%¹05%with oil samples.For deoxynivalenol,the method can achieve the lowest detection limit of 0.069ng/mL,the lowest limit of quantification of 0.91ng/mL,the intra-and inter-assay variation both below 11%,and the recovery rate is within 87.8%¹06.0%with oil samples.In summary,the luminescent oxygen channeling immunoassay technology can meet the rapid and high-throughput detection requirements of mycotoxins in agricultural products and has broad application prospects in the field of rapid detection of mycotoxins.