Establishment of a blocking ELISA using a newly generated monoclonal antibody against Salmonella flagellin for detecting antibodies in infected chickens.

backgroundSalmonella is an important zoonotic pathogen that is widely distributed in every link between livestock and poultry production. Salmonella contamination not only affects the healthy development of the livestock industry but also poses a serious threat to public health, as it can infect humans through farming, processing, and consumption of meat and egg products. Therefore, establishing detection methods capable of simultaneously identifying different Salmonella serotypes is crucial.

resultsThis study focused on using Salmonella flagellin as an immunogen to screen and prepare monoclonal antibodies, ultimately establishing a blocking ELISA (bELISA). In this study, flagellin from S. Enteritidis was extracted by acid lysis and used to immunize BALB/c mice. Cell fusion was performed following three immunizations, resulting in the generation of six hybridoma cell lines that stably secreted monoclonal antibodies. Monoclonal antibody (MAb) of 1F6 exhibited excellent reactivity with Salmonella flagellin, demonstrating an affinity of 1.11 × 10

conclusionsThe bELISA method was developed by utilizing Salmonella flagellin as an antigen and exhibited excellent stability, strong specificity, and sensitivity, making it suitable for broad-spectrum screening of Salmonella in infected chicken.