Abstract: [Objective] To establish a rapid detection method for aflatoxin B₁ （AFB₁）. [Method] The acridinium ester （AE） and AFB₁ monoclonal antibody were chemically synthesized in vitro, and the AE labeled AFB₁ monoclonal antibody （AFB₁-AE） with different molar ratios of AE and AFB₁ monoclonal antibody of 5∶1, 10∶1, 15∶1, 20∶1 and 25∶1 was synthesized, respectively; the detection effects of AFB₁-AE conjugates with different molar ratios were analyzed, and the conjugate with molar ratio of 25∶1 of AE and AFB₁ monoclonal antibody was finally selected to establish chemiluminescence immunoassay （CLIA）. [Result] The corresponding regression line equation was obtained: y=-0.245Ln（x） + 1.578 1, R²=0.985 7, IC₅₀=81.48 pg/mL; the sample recovery experiment was carried out, and the standard recovery rate was 88.4%-106.0%, the coefficient of variation was 0.92%-2.10%. [Conclusion] In vitro cross-linking of AE with AFB₁ monoclonal antibody can be used to establish a new chemiluminescence immunoassay method.

Key words: aflatoxin B₁, acridinium ester, chemiluminescence immunoassay, coupling