Novel dual immunochromatographic test strip based on double antibodies and biotin-streptavidin system for simultaneous sensitive detection of aflatoxin M1 and ochratoxin A in milk

The coexistence of mycotoxins in agricultural products poses a serious threat to food safety. This study developed a dual immunochromatographic test strips (DICTS) method based on double antibodies labeled with time-resolved fluorescent microspheres (TRFM) to realize simultaneous rapid detection of aflatoxin M1 (AFM1) and ochratoxin A (OTA) in milk. As bridge antibody, the polyclonal antibody (pAb) was first conjugated with the TRFM and then with the monoclonal antibody (mAb). Meanwhile, a biotin-streptavidin system was introduced to replace the traditional goat anti-mouse Immunoglobulin G, thus providing a stable signal on the control line. After optimization, the detection limit of AFM1 and OTA in milk was respectively 0.018 and 0.036 ng/mL. The recoveries of intraassay and interassay experiments ranged from 89.65% to 103.99%. The accuracy, repeatability, and specificity of the developed TRFM-DICTS were estimated. The results of TRFM-DICTS showed a high consistency with those of the ultrahigh-performance liquid chromatography-tandem mass spectrometry.