Development of a low-cost paper-based ELISA method for rapid Escherichia coli O157:H7 detection

Escherichia coli O157: H7 (E. coli O157: H7) has become one of the most dangerous foodborne pathogenic bacteria around the world. Currently, because of the tedious, high-cost and stringent laboratory conditions required, the conventional E. coli O157: H7 detection methods, such as culture-based methods and polymerase chain reaction (PCR), have much limitation. Thus, we developed a novel paper-based enzyme-linked immunosorbent assay (p-ELISA) with shorter operation duration, lower cost, relatively higher sensitivity and wider application. This method required less than 3 h and 5 μL of sample to complete the detection. The limit of detection (LOD) for E. coli O157: H7 reached 1 × 10⁴ CFU/mL with high specificity. To be more suitable for on-site testing, the readout could be rapidly obtained without any expensive instruments. In this study, we chose E. coli O157:H7 as the representative, and our method could provide a platform for determination of other pathogenic bacteria.