Sensitive detection and point-of-care test of bacterial pathogens is of great significance in safeguarding the public health worldwide. Inspired by the characteristics of horseradish peroxidase (HRP), we synthesized a hybrid nanoflower with peroxidase-like activity via a three-component self-assembled strategy. Interestingly, the prepared nanozyme not only could act as an alternative to HRP for colorimetric biosensing, but also function as a unique signal probe that could be recognized by a pregnancy test strip. By combining the bifunctional properties of hybrid nanoflower, isothermal amplification of LAMP, and the specific recognition and non-specific cleavage properties of CRISPR/Cas12a system, the dual-readout CRISPR/Cas12a biosensor was developed for sensitive and rapid detection of Salmonella enterica. Moreover, this platform in the detection of Salmonella enterica had limits of detection of 1 cfu/mL (colorimetric assay) in the linear range of 101-108 cfu/mL and 102 cfu/mL (lateral flow assay) in the linear range of 102-108 cfu/mL, respectively. Furthermore, the developed biosensor exhibited good recoveries in the spiked samples (lake water and milk) with varying concentrations of Salmonella enterica. This work provides new insights for the design of multifunctional nanozyme and the development of innovative dual-readout CRISPR/Cas system-based biosensing platform for the detection of pathogens.

A novel bifunctional MOF-encapsulated cobalt-doped carbon dots nanozyme (Co-CD/PMOF) with excellent peroxidase-mimic catalytic activity and fluorescence property was synthesized and employed to fabricate a chemiluminescence/fluorescence (CL/FL) dual-mode immunosensor for AFB1 detection. Co-CD/PMOF could catalyze the luminol/H2O2 system to generate robust and long-lasting CL signals due to the slow diffusion effect and continuous generation of •OH, O2•-, and 1O2 species. Differing from traditional flash-type CL emissions, this glow-type CL emission is helpful to fabricate a sensitive and accurate CL sensing platform. Then the CL/FL dual-mode detection of AFB1 was developed using antibody-functionalized Co-CD/PMOF as the signal-amplifying nanoprobe. The CL mode assay based on indirect competitive immune principle was carried out on a chemiluminescence optical fiber platform, where the AFB1-OVA-functionalized optical fiber probe was employed for biorecognition, separation, and signal conducting. The AFB1 detection range and LOD were 0.63-69.36 ng/mL and 0.217 ng/mL, respectively. Using AFB1 antibody-functionalized immunomagnetic beads for capturing and separation, the FL mode detection of AFB1 was established based on the sandwich immune principle. A linear range of 0.54-51.91 ng/mL and a LOD of 0.027 ng/mL were obtained. This work designed a sensitive, rapid, and reliable nanozyme-powered dual-mode assay strategy and provided technical support in the field of environmental monitoring and food safety.

Alzheimer\'s disease (AD) is one of the most common age-associated brain diseases and is induced by the accumulation of amyloid beta (Aβ) and oxidative stress. Many studies have focused on eliminating Aβ by nanoparticle affinity; however, nanoparticles are taken up mainly by microglia rather than neurons, leading poor control of AD. Herein, mitochondria-targeted nanozymes known as (3-carboxypropyl)triphenyl-phosphonium bromide-conjugated 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000]-functionalized molybdenum disulfide quantum dots (TPP-MoS2 QDs) were designed. TPP-MoS2 QDs mitigate Aβ aggregate-mediated neurotoxicity and eliminate Aβ aggregates in AD mice by switching microglia from the proinflammatory M1 phenotype to the anti-inflammatory M2 phenotype. TPP-MoS2 QDs cross the blood-brain barrier, escape from lysosomes, target mitochondria and exhibit the comprehensive activity of a bifunctional nanozyme, thus preventing spontaneous neuroinflammation by regulating the proinflammatory substances interleukin-1β, interleukin-6 and tumor necrosis factors as well as the anti-inflammatory substance transforming growth factor-β. In contrast to the low efficacy of eliminating Aβ by nanoparticle affinity, the present study provides a new pathway to mitigate AD pathology through mitochondria-targeted nanozymes and M1/M2 microglial polarization.