ABSTRACT
A new catechol-substituted monostyryl boradiazaindacene (BODIPY)-based lanthanide-free colorimetric and fluorogenic probe was developed for the sequential detection of Cu2+ ions and dipicolinic acid (DPA), a distinctive biomarker of bacterial endospores, with high sensitivity and selectivity. In the presence of Cu2+ ions, the blue solution of the probe changes to cyan and the fluorescence is quenched, however, the cyan color changes to blue immediately and the fluorescence is restored on contact with DPA, resulting from competitive binding of DPA that interact with Cu2+ ions. A practical application by using Geobacillus stearothermophilus spores was further studied and as low as 1.0 x 105 spores were detected.
Subject(s)
Biomarkers/analysis , Boron Compounds/chemistry , Copper/analysis , Fluorescent Dyes/chemistry , Picolinic Acids/analysis , Spores, Bacterial/chemistry , Color , Colorimetry , Geobacillus stearothermophilus/chemistry , Geobacillus stearothermophilus/metabolism , Indicators and Reagents , Lanthanoid Series Elements , Sensitivity and SpecificityABSTRACT
Food safety is a major issue to protect public health and a key challenge is to find detection methods for identification of hazards in food. Food borne infections affects millions of people each year and among pathogens, Salmonella Enteritidis is most widely found bacteria causing food borne diseases. Therefore, simple, rapid, and specific detection methods are needed for food safety. In this study, we demonstrated the selection of DNA aptamers with high affinity and specificity against S. Enteritidis via Cell Systematic Evolution of Ligands by Exponential Enrichment (Cell-SELEX) and development of sandwich type aptamer-based colorimetric platforms for its detection. Two highly specific aptamers, crn-1 and crn-2, were developed through 12 rounds of selection with Kd of 0.971µM and 0.309µM, respectively. Both aptamers were used to construct sandwich type capillary detection platforms. With the detection limit of 103 CFU/mL, crn-1 and crn-2 based platforms detected target bacteria specifically based on color change. This platform is also suitable for detection of S. Enteritidis in complex food matrix. Thus, this is the first to demonstrate use of Salmonella aptamers for development of the colorimetric aptamer-based detection platform in its identification and detection with naked eye in point-of-care.