A PCR-free genosensing platform for detection of Shigella dysenteriae in human plasma samples by porous and honeycomb-like biochar decorated with ultrathin flower-like MoS2 nanosheets incorporated with Au nanoparticles

Abstract

Shigella dysenteriae, a gram-negative bacterium, which results in the most infectious of bacterial shigellosis and dysenteries. In this study, an innovative gene detection platform based on label-free DNA sequences was developed to detect Shigella dysenteriae in human plasma samples. The porous and honeycomb-like structure of biochar (BC) was first synthesized through a pyrolysis process. Then, the produced biochar was effectively decorated with flower-like MoS2 nanosheets (MoS2/BC). The resulting nanocomposite was incorporated with Au nanoparticles (AuNPs) by applying chronoamperometry technique, and then the subsequent product including MoS2 nanosheets, biochar and AuNPs were immobilized on the Au electrode surface and used for modifier agent in electrochemical bio-assays. Structural and morphological study of the synthesized compounds were investigated using various characterization methods such as FE-SEM, TEM, EDS, FTIR, and XRD. Various electrochemical techniques including cyclic voltammetry (CV) and Differential pulse anodic stripping voltammetry (DPASV) have been used to investigate the applicability of the fabricated genosensing bio-assay. Under optimal conditions, LOD and LOQ were calculated 9.14 fM and 0.018 pM respectively. In addition, a linear range from 0.01 to 100 pM was obtained for single stranded-target DNA (ss-tDNA), with R-2 of 0.9992. The recoveries ranged from 98.0 to 101.3%. The fabricated bio-detection assay demonstrated high selectivity for 1, 2, and 3 base mismatch sequences. In addition, a negative control of the gene detection platform which was performed to study selectivity was provided by ss-tDNA from Haemophilus influenzae, and Salmonella typhimurium. Moreover, it is important to mention that the organized bioassay is simply reusable and reproducible with the RSD% (relative standard deviation) < 5 to next detection assays.