Targeting the cell wall: Preparation of monoclonal antibody for accurate identification of Alicyclobacillus acidoterrestris in apple juice

Alicyclobacillus acidoterrestris represents the predominant microorganism implicated in apple juice spoilage. Most of existing detection methods for this microorganism necessitate advanced equipment and skilled technicians, both of which are unsuitable for practical application. In this study, an effective approach for the identification of A. acidoterrestris was developed based on an indirect sandwich enzyme-linked immunosorbent assay (is-ELISA). This method utilizes the species-specific monoclonal antibody (mAb) targeting a cell wall protein of A. acidoterrestris. The is-ELISA was highly sensitive with a limit of detection (LOD) of 5.4 × 102 CFU/mL. Furthermore, the assay is capable of accurately differentiating between A. acidoterrestris and other species of genus Alicyclobacillus. The applicability of is-ELISA was also evaluated in artificially contaminated apple juice. The convenience, simplicity, sensitivity and specificity of this assay represents a robust method for the detection of A. acidoterrestris and the resultant prevention of the spoilage of fruit juices. [Display omitted] •A novel cell wall protein of Alicyclobacillus acidoterrestris was obtained for the first time.•A monoclonal antibody only specific for A. acidoterrestris was prepared for the first time.•An indirect sandwich ELISA was developed for accurate detection A. acidoterrestris with high sensitivity.•Positive result can be obtained after 22 h enrichment in apple juice contaminated with 1 CFU/mL of A. acidoterrestris.