Viable E. coli O157:H7 detection based on Föster resonance energy transfer (FRET) system using FITC and TRITC conjugates as molecular probes
Year: 2025
Type: article
Abstract: Improved rapid and sensitive biosensors for detecting foodborne pathogens in the environment are urgently needed to curtail disease outbreaks. This study aims to develop and optimize a fast and sensitive biosensor for detecting E. coli O157 using an advanced Förster Resonance Energy Transfer (FRET) technology. Building on recent FRET technological advancements, we intend to measure the degree of interaction between primary antibodies labeled with fluorescein isothiocyanate (FITC) and secondary antibodies labeled with tetramethyl rhodamine isothiocyanate (TRITC) in a sandwich format. This study assesses the detection of E. coli O157 at various pH levels in contaminated lettuce juice. A detection range of 10 2 to 10 4 CFU/mL after a 4-hour pre-enrichment at 37°C enabled sensitive quantification. In this FRET-based method, FITC-IgG was used as the donor and TRITC-IgY as the acceptor. Samples were stimulated at 350 nm, and emissions were measured from 500 to 680 nm, capturing a wide range of fluorescence signals. FRET microscopic fluorescence imaging was used to determine how pH affects the concentration of live E. coli O157. The results showed that pH change has a considerable impact on detection. The approach illustrates FRET’s effectiveness in identifying live pathogens under diverse settings, indicating its underlying relevance in pathogen identification.
Source: MethodsX
Language: English
Subfield: Biomedical Engineering
Field: Engineering
Domain: Physical Sciences
Open Access status: gold
Estimated APC paid: $1,260
OpenAlex ID: W4411079192