Since the ferromagnetic (Fe3O4) nanoparticles were firstly reported to exert enzyme-like activity in 2007, extensive research progress in nanozymes has been made with deep investigation of diverse nanozymes and rapid development of related nanotechnologies. As promising alternatives for natural enzymes, nanozymes have broadened the way toward clinical medicine, food safety, environmental monitoring, and chemical production. The past decade has witnessed the rapid development of metal- and metal oxide-based nanozymes owing to their remarkable physicochemical properties in parallel with low cost, high stability, and easy storage. It is widely known that the deep study of catalytic activities and mechanism sheds significant influence on the applications of nanozymes. This review digs into the characteristics and intrinsic properties of metal- and metal oxide-based nanozymes, especially emphasizing their catalytic mechanism and recent applications in biological analysis, relieving inflammation, antibacterial, and cancer therapy. We also conclude the present challenges and provide insights into the future research of nanozymes constituted of metal and metal oxide nanomaterials.

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A Review on Metal- and Metal Oxide-Based Nanozymes: Properties, Mechanisms, and Applications

Recent advances in gold nanoparticles-based biosensors for food safety detection

The pervasive spread of infectious diseases and pandemics, such as the 2019 coronavirus disease (COVID-19), are becoming increasingly serious and urgent threats to human health. Preventing the spread of such diseases prioritizes the development of sensing devices that can rapidly, selectively, and reliably detect pathogens at minimal cost. Paper-based analytical devices (PADs) are promising tools that satisfy those criteria. Numerous paper-based biosensors have been established that rival conventional pathogen detection methods. Among them, colorimetric strategies are promising since results can be interpreted by eye, and are simple to operate, which is advantageous for point-of-care testing (POCT). Particularly, the application of nanomaterials on paper-based biosensors has become important as these materials are capable of converting signals from pathogens through unique mechanisms to yield an amplified colorimetric readout. To highlight the research progress on using nanomaterials in colorimetric paper-based biosensor for pathogen detection, we discuss the sensing mechanisms of how they work, structural and analytical characteristics of the devices, and representative recent applications. Current challenges and future directions of using PADs and nanomaterial-mediated strategies are also discussed.

Nanomaterial-mediated paper-based biosensors for colorimetric pathogen detection.

Since its first discovery in December 2019, the global coronavirus disease 2019 (COVID-19) pandemic caused by the novel coronavirus (SARS-CoV-2) has been posing a serious threat to human life and health. Diagnostic testing is critical for the control and management of the COVID-19 pandemic. In particular, diagnostic testing at the point of care (POC) has been widely accepted as part of the post restriction COVID-19 control strategy. Lateral flow assay (LFA) is a popular POC diagnostic platform that plays an important role in controlling the COVID-19 pandemic in industrialized countries and resource-limited settings. Numerous pioneering studies on the design and development of diverse LFA-based diagnostic technologies for the rapid diagnosis of COVID-19 have been done and reported by researchers. Hundreds of LFA-based diagnostic prototypes have sprung up, some of which have been developed into commercial test kits for the rapid diagnosis of COVID-19. In this review, we summarize the crucial role of rapid diagnostic tests using LFA in targeting SARS-CoV-2-specific RNA, antibodies, antigens, and whole virus. Then, we discuss the design principle and working mechanisms of these available LFA methods, emphasizing their clinical diagnostic efficiency. Ultimately, we elaborate the challenges of current LFA diagnostics for COVID-19 and highlight the need for continuous improvement in rapid diagnostic tests.

Point-of-care COVID-19 diagnostics powered by lateral flow assay.

The conventional colloidal gold immunochromatographic assay (AuNP-ICA) cannot meet the requirements for the rapid and sensitive detection of Escherichia coli (E. coli) O157:H7 because of its poor sensitivity. Herein, a novel two-step cascade signal amplification strategy that integrates in situ gold growth and nanozyme-mediated catalytic deposition was proposed to enhance the detection sensitivity of conventional AuNP-ICA dramatically. The enhanced strip displayed ultrahigh sensitivity in E. coli O157:H7 detection and had a detection limit of 1.25 × 101 CFU/mL, which is approximately 400-fold lower than that of traditional AuNP-ICA (5 × 103 CFU/mL). The amplified strip had no background signal in the T-line zone in the absence of E. coli O157:H7 even after one round of cascade signal amplification. The enhanced strip demonstrated excellent selectivity against E. coli O157:H7 with a negligible cross-reaction to nine other common pathogens. Intra-assays and interassays showed that the improved strip has acceptable accuracy and precision for determining E. coli O157:H7. The average recoveries in a real milk sample ranged from 87.33 to 112.15%, and the coefficients of variation were less than 10%. The enhanced strip also showed great potential in detecting a single E. coli O157:H7 cell in a 75 μL solution.

Dramatically Enhanced Immunochromatographic Assay Using Cascade Signal Amplification for Ultrasensitive Detection of Escherichia coli O157:H7 in Milk.

Gold nanoflowers (GNFs) exhibit stronger light scattering ability than gold nanospheres (GNSs) with the same diameter, thereby contributing to enhancing the sensitivity of the scattering-based sensing method. However, the application of GNFs in biosensors based on dynamic light scattering (DLS) has not been yet reported. Herein, we describe for the first time an improved no-wash immunosensor based on dynamic light scattering for the detection of Escherichia coli O157:H7 (E. coli O157:H7) in milk using GNFs for sensitive signal transduction. To achieve this goal, a thiolated amphiphilic carboxyl ligand was introduced to modify the GNF surface and improve solution stability and antibody functionalization. Several key factors that affect the detection sensitivity of our developed GNF_DLS immunosensor were systematically investigated. Under the optimal conditions, our proposed GNF_DLS immunosensor provided an excellent linear detection for E. coli O157:H7 within the range from 6 × 100 to 6 × 104 colony-forming units (CFU)/mL, with a limit of detection of 2.7 CFU/mL. Combined with our previously reported two-step large-volume immunomagnetic separation (IMS) method, the designed GNF_DLS immunosensor can sensitively, selectively, and accurately detect the presence of E. coli O157:H7 in pasteurized milk. The potential of our GNF_DLS method for monitoring the presence of a single bacterial cell in 1 mL of sample solution was also demonstrated. Overall, the developed GNF_DLS immunosensor can be used for the rapid and high-sensitivity determination of pathogenic bacteria and can be extended for the ultrasensitive no-wash detection of other trace analytes.

Gold Nanoflower-Enhanced Dynamic Light Scattering Immunosensor for the Ultrasensitive No-Wash Detection of Escherichia coli O157:H7 in Milk.

Herein, we report a novel multiplex fluorescent competitive immunosensor using self-luminous bacteria as a bifunctional probe for the simultaneous detection and quantification of multiple pathogens. The self-luminous bacteria are prepared through the facile one-step co-culture of bacteria and organic fluorescent dyes. Cronobacter muytjensii, Listeria monocytogenes, and Escherichia coli O157:H7 are fluorescently labelled with 7-hydroxycoumarin, fluorescein isothiocyanate, and rhodamine B, respectively, due to the dyes small spectral overlap in excitation/emission. The obtained fluorescent bacteria retain specific immunorecognition activity, but exhibit excellent fluorescent sensing ability. By using these fluorescent bacteria as bifunctional probes for competitive recognition and optical sensing, we achieved the sensitive and simultaneous quantitative determination of three pathogens with sensitivities of below 8.6 × 103 CFU/mL. The sensitivities can be significantly enhanced to 1 CFU/mL in real milk by integrating with a pre-incubation step, showing great potential for the ultrasensitive determination of trace bacteria and even single colony with the overall detection time of less than 10 h. In summary, this work provides an alternative multiplexing analytical platform as a powerful and rapid diagnostic tool for monitoring foodborne pathogen contamination.

Jinmei Fu

Papers

Dramatically Enhanced Immunochromatographic Assay Using Cascade Signal Amplification for Ultrasensitive Detection of Escherichia coli O157:H7 in Milk.

Gold Nanoflower-Enhanced Dynamic Light Scattering Immunosensor for the Ultrasensitive No-Wash Detection of Escherichia coli O157:H7 in Milk.

A self-luminous bifunctional bacteria directed fluorescent immunosensor for the simultaneous detection and quantification of three pathogens in milk