Nanobody-based immunomagnetic separation platform for rapid isolation and detection of Salmonella enteritidis in food samples

Abstract

Rapid separation and identification of Salmonella enteritidis (S. enteritidis) in food is of great importance to prevent outbreaks of foodborne diseases. Herein, by using O and H antigens as targets, an epitope-based bio-panning strategy was applied to isolate specific nanobodies towards S. enteritidis. This method constitutes an efficient way to obtain specific antibody fragments and test pairwise nanobodies. On this basis, a double nanobody-based sandwich enzyme-linked immunosorbent assay (ELISA) coupled with immunomagnetic separation (IMS) was developed to rapid enrich and detect S. enteritidis in food. The detection limit of the IMS-ELISA was 3.2 × 10³ CFU/mL. In addition, 1 CFU of S. enteritidis in food samples can be detected after 4-h cultivation, which was shortened by 2 h after IMS. The IMS-ELISA strategy could avoid matrix interference and shorten the enrichment culture time, which has great potential for application in monitoring bacterial contamination in food.

Keywords: ELISA; Immunomagnetic separation; Rapid detection; Salmonella; Single domain antibody.