A novel lateral flow immunoassay strip based on a label-free magnetic Fe3O4@UiO-66-NH2 nanocomposite for rapid detection of Listeria monocytogenes

Abstract

Listeria monocytogenes (L. monocytogenes) is one of the most lethal pathogenic bacteria. Although the traditional microbial culture method has high sensitivity and selectivity for the diagnosis of L. monocytogenes, it is time-consuming and not suitable for on-site detection. A rapid, convenient and visualized on-site detection method is particularly needed. In this work, Fe₃O₄@UiO-66-NH₂ was prepared for both magnetic separation and lateral flow immunoassay (LFIA) for the detection of L. monocytogenes by taking advantage of the easy separation of the magnetic core Fe₃O₄ and the high surface area of the outer layer UiO-66-NH₂. Fe₃O₄@UiO-66-NH₂ with a high surface area and good water-dispersibility and optical properties was synthesized by a simple hydrothermal process. It could directly adsorb on the surface of target bacteria and form Fe₃O₄@UiO-66-NH₂-bacteria conjugates, without the labeling of an antibody. After magnetic separation and concentration, the Fe₃O₄@UiO-66-NH₂-bacteria conjugates were detected by the antibody on the test line of the LFIA strip, resulting in a visible orange band. The capture efficiency and LFIA detection of Fe₃O₄@UiO-66-NH₂ were optimized in this study. Under the optimal conditions, a good linear correlation between the test line intensity and the concentration of L. monocytogenes was obtained in the range of 10⁵–10⁸ CFU mL⁻¹, and the limit of detection was 2.2 × 10⁶ CFU mL⁻¹ by the naked eye. The Fe₃O₄@UiO-66-NH₂-based LFIA strip showed strong specificity for L. monocytogenes, and the detection took 45 min without culture enrichment. Therefore, the proposed Fe₃O₄@UiO-66-NH₂-based strip showed the advantages of simple synthesis, being label-free, low cost, good selectivity and convenience.