Bimetallic Fe/Mn MOFs/MβCD/AuNPs stabilized on MWCNTs for developing a label-free DNA-based genosensing bio-assay applied in the determination of Salmonella typhimurium in milk samples

Abstract

Monitoring of pathogenic bacteria plays a vital role in precluding foodborne disease outbreaks. In this research work, a genosensor based on innovative label-free DNA was developed for the detection of Salmonella. typhimurium (S. typhimurium) in the milk samples. To realize this objective, bimetallic Fe/Mn MOF is synthesized and mixed with methyl-β-cyclodextrin (MβCD) and AuNPs which are then stabilized on multi-walled carbon nanotubes (MWCNTs), and the obtained nanocomposite is immobilized on the Au electrode surface. Different characterization methods such as FE-SEM, TEM, EDS, FTIR, and XRD were used for investigating the particle size and morphological features. Electrochemical and impedimetric techniques were used for exploring the applicability of the fabricated genosensor. Under optimal circumstances, LOD and LOQ have acquired at 0.07 pM and 0.21 pM. Moreover, an extensive linear range of 1 pM-1 μM was resulted for ss-tDNA (single-stranded target DNA), R² obtained 0.9991. The recoveries were obtained 95.6-104%. Great selectivity against one, two, and three-base mismatched sequences was also shown for fabricated biosensing assay. Furthermore, negative genosensing assay control for investigating selectivity was provided by the ss-tDNAs of Haemophilusinfluenzae and Shigella dysenteriae bacteria. Well-fabricated genosensing bio-assay represents better performance, great specificity, high sensitivity, increased active sites, and finally results in an increase in the electron transfer rate. It is to be noted that the organized genosensing bio-assay is capable of being re-used and re-generated in a straightforward manner to estimate the hybridization process.

Keywords: AuNPs; Bimetallic Fe/Mn MOFs; DNA hybridization; Genosensor; Multi-walled carbon nanotubes; Salmonella typhimurium.