ABSTRACT
Simultaneous detection of multiple foodborne pathogens is of great importance for ensuring food safety. Herein, we present a sensitive dual-channel electrochemical biosensor based on copper metal organic frameworks (CuMOF) and lead metal organic framework (PbMOF) for simultaneous detection of Salmonella typhimurium (S. typhimurium) and Listeria monocytogenes (L. monocytogenes). The MOF-based nanotags were prepared by functionalizing gold nanoparticles loaded CuMOF (Au@CuMOF) and PbMOF (Au@PbMOF) with signal DNA sequences 1 (sDNA1) and sDNA2, respectively. By selecting invA of S. typhimurium and inlA gene of L. monocytogenes as targe sequences, a sandwich-typed dual-channel biosensor was developed on glassy carbon electrodes (GCE) through hybridization reactions. The sensitive detection of S. typhimurium and L. monocytogenes was achieved by the direct differential pulse voltametric (DPV) signals of Cu2+ and Pb2+. Under optimal conditions, channel 1 of the biosensor showed linear range for invA gene of S. typhimurium in 1 × 10-14-1 × 10-8 M with low detection limit (LOD) of 3.42 × 10-16 M (S/N = 3), and channel 2 of the biosensor showed linear range for inlA gene of L. monocytogenes in 1 × 10-13-1 × 10-8 M with LOD of 6.11 × 10-15 M (S/N = 3). The dual-channel biosensor showed good selectivity which were used to detect S. typhimurium with linear range of 5-1.0 × 104 CFU mL-1 (LOD of 2.33 CFU mL-1), and L. monocytogenes with linear range of 10 - 1.0 × 104 CFU mL-1 (LOD of 6.61 CFU mL-1).
Subject(s)
Biosensing Techniques , Metal Nanoparticles , Metal-Organic Frameworks , Gold , Salmonella typhimurium , Limit of DetectionABSTRACT
The present work developed an electrochemical genosensor for the detection of virulence outer membrane protein A (ompA, tDNA) gene of Cronobacter sakazakii (C. sakazakii) by exploiting the excellent glucose-oxidase-mimicking activity of copper Metal-organic frameworks (Cu-MOF) doped with gold nanoparticle (AuNPs). The signal nanotags of signal probes (sDNA) that biofunctionalized AuNPs@Cu-MOF (sDNA-AuNPs@Cu-MOF) were designed using an Au-S bond. The biosensor was prepared by immobilization capture probes (cDNA) onto an electrodeposited AuNPs-modified glassy carbon electrode (GCE). AuNPs@Cu-MOF was introduced onto the surface of the GCE via a hybridization reaction between cDNA and tDNA, as well as tDNA and sDNA. Due to the enhanced oxidase-mimicking activity of AuNPs@Cu-MOF to glucose, the biosensor gave a linear range of 1.0 × 10-15 to 1.0 × 10-9 mol L-1 to tDNA with a detection limit (LOD) of 0.42 fmol L-1 under optimized conditions using differential pulse voltammetry measurement (DPV). It can be applied in the direct detection of ompA gene segments in total DNA extracts from C. sakazakii with a broad linear range of 5.4-5.4 × 105 CFU mL-1 and a LOD of 0.35 CFU mL-1. The biosensor showed good selectivity, fabricating reproducibility and storage stability, and can be used for the detection of ompA gene segments in real samples with recovery between 87.5% and 107.3%.
