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An Improved Chemiluminescence Immunoassay for the Ultrasensitive Detection of Aflatoxin B1

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Abstract

An indirect competitive chemiluminescence (CL) immunoassay based on Fe3O4@SiO2@Au magnetic nanoparticles (Au-SCMPs) has been optimized and developed for the detection of aflatoxin B1 (AFB1). To improve the detection sensitivity and efficiency, this method combines 2′,6′-dimethylcarbonylphenyl-10-sulfopropyl acridinium-9-carboxylate 4′-NHS ester (NSP-DMAE-NHS) as a new kind of high efficient luminescence reagent with a simplified separation procedure based on the optimized Au-SCMPs. Superparamagnetic nanoparticles were coated with different sizes of Au colloids (18, 30, and 50 nm), and their capacities in immobilization of bovine serum albumin (BSA) were investigated. The BSA-AFB1 conjugate (BSA-AFB1) was immobilized on the optimized Au-SCMPs and competed with the free AFB1 for specially binding the NSP-DAME-NHS-labeled anti-AFB1 antibody (mAb-AFB1). After the indirect competitive immunoreactions and magnetic separation, the CL of the nanoparticles was measured by a homemade luminescent measurement system. Under the optimum conditions, the IC50 value was 0.07 ng mL−1 and the limit of detection (LOD) of AFB1 was 0.01 ng mL−1, respectively. The result shows a much lower IC50 and LOD than that typically achieved by ELISA. This proposed immunoassay method was rapid, low-cost, and suitable for the detection of AFB1.

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Authors and Affiliations

  1. Physics Department, School of Science, Xi’an Jiaotong University, Xi’an, People’s Republic of China

    Junfeng Li, Xiangyi Fang & Peng Tang

  2. Department of Clinical Laboratory, The First Affiliated Hospital of Medical College, Xi’an Jiaotong University, Xi’an, People’s Republic of China

    Yucong Yang & Xiaoli Cheng

Authors
  1. Junfeng Li
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  2. Xiangyi Fang
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  3. Yucong Yang
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  4. Xiaoli Cheng
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  5. Peng Tang
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Correspondence to Xiangyi Fang.

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Funding

This work was supported by the National Natural Science Foundation of China (NSFC, Grant No. 81371642) and the Fundamental Research Funds for the Central Universities of China (XKJC2013005).

Conflict of Interest

Junfeng Li declares that he has no conflict of interest. Xiangyi Fang declares that he has no conflict of interest. Yucong Yang declares that she has no conflict of interest. Xiaoli Cheng declares that she has no conflict of interest. Peng Tang declares that he has no conflict of interest.

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Li, J., Fang, X., Yang, Y. et al. An Improved Chemiluminescence Immunoassay for the Ultrasensitive Detection of Aflatoxin B1 . Food Anal. Methods 9, 3080–3086 (2016). https://doi.org/10.1007/s12161-016-0499-1

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  • DOI: https://doi.org/10.1007/s12161-016-0499-1

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