A rapid and ultrasensitive paper sensor for Haemolysin BL detection

Bacillus cereus is a foodborne opportunistic pathogen commonly found in humans and animals. It produces various toxins, causing frequent food safety incidents. Therefore, establishing a fast and accurate method for detecting B. cereus enterotoxin is crucial for disease diagnosis and food safety. In this study, Haemolysin BL comprising Hbl B and L2 was obtained from a prokaryotic expression system and then used to immunize mice for antibody preparation. Paired antibodies 2A10-5C7 against Hbl B and 1E2-10A4 against Hbl L2 were screened using the chessboard method and then used to construct a double-antibody sandwich detection method and a lateral flow immunochromatographic assay (LFIA) to quantify the concentrations of Hbl B and L2 in PBS and milk. The limits of detection for Hbl B and L2 in milk were 0.74 ng mL −1 and 1 ng mL −1 with detection ranges of 1.48-645.5 ng mL −1 and 2.33-391.5 ng mL −1 . The spiked recoveries ranged from 82.2% to 105.67% and there was no cross-reactivity with common microbial toxins. The established LFIA was low in cost and rapid and was comparable with commercially available detection kits for food samples. LFIA has been successfully applied to actual sample detection for Hbl B and L2, with an LOD of 1 ng mL −1 .