Universal and reusable hapten/antibody-mediated portable optofluidic immunosensing platform for rapid on-site detection of pathogens

Abstract

A universal and reusable hapten–antibody-mediated portable optofluidic immunosensing platform (OIP) was developed for rapid on-site detection of pathogens. By using Escherichia coli O157:H7 (E. coli O157:H7) and bisphenol A-Bovine serum albumin (BPA-BSA)/anti-BPA antibody as a model pathogen and a mediated hapten–antibody, respectively, a novel immunoassay mechanism was proposed to detect pathogens. The BPA-BSA-modified immunosensor and E. coli O157:H7 were initially saturated with anti-BPA antibodies (mouse IgG) and anti-E. coli O157:H7 antibodies (mouse IgG), respectively. Then, the fluorescence-labeled secondary antibodies (goat anti-mouse IgG antibody) were incubated with E. coli O157:H7 with their antibodies. Next, the mixture was introduced into the immunosensor surface bound to the anti-BPA antibodies. A high concentration of E. coli O157:H7 in the sample reduced the number of fluorescence-labeled secondary antibodies bound to the immunosensor surface, thus resulting in the detection of low fluorescence signals. Under optimized conditions, the hapten–antibody-mediated OIP system exhibited a detection limit of 8 cfu/mL E. coli O157:H7 after concentrating 100 times by using centrifugation, and a test cycle, including prereaction, detection, and regeneration, was less than 1 h. The robustness of the hapten–carrier protein-modified immunosensor surface allowed multiple pathogen immunoassays. The proposed strategy demonstrated good recovery, precision, and accuracy through the evaluation of the spiked water samples. We expect that the new platform can be readily used for the detection of other pathogens in a variety of application fields ranging from environmental monitoring and food safety to medical diagnosis.