Colorimetric Biosensor Research Articles

Covalent copper-organic frameworks with light-activable catalytic centers as smart artificial enzymes for highly sensitive and wide-range biocatalytic diagnosis

As a popular sensing method, colorimetric biosensing has the advantages of convenient operation, low cost, fast response, and easy readout. However, due to the complex structure of nanomaterials, low catalytic activity, and unclear catalytic mechanisms, constructing efficient, stable, and specific artificial biocatalysts for colorimetric biosensing is still a challenge. In this study, to propose a new colorimetric biosensing system, we synthesize three different types of copper cyclic trinuclear unit-based 2D copper-organic frameworks (CuOFs) with covalent structures and light-activable catalytic centers to serve as smart artificial enzymes for highly sensitive and wide-range biocatalytic diagnosis. By varying the light absorption centers, CuOFs with different band structures, light absorption capacity, photoelectron transfer efficiency, and electron-hole pair separation efficiency can be obtained. Of the three covalent CuOFs, tris(4-aminophenyl)amine (Tpa)-Cu3 exhibited the widest light absorption range, the narrowest band gap, the smallest electron transfer barrier, and the best-photoinduced charge separation efficiency, which was conducive to improving the light capture ability and eventually improving the photocatalytic properties. Further tests demonstrated that Tpa-Cu3 exhibited the best peroxidase-like activity under visible light irradiation, which could effectively diagnose the glucose and common antioxidants with a wider detection range and lower detection limit than many previously reported biocatalytic materials. We expect this design will provide new insights into enzyme-like organic framework materials and unique systems for high-sensitive and wide-range colorimetric detection for future biocatalytic applications.

Assembly of Peptide with Dye Molecules for the Fabrication of Colorimetric Biosensor with Application To Diagnose HER2-Positive Breast Cancer

Assembly of Peptide with Dye Molecules for the Fabrication of Colorimetric Biosensor with Application To Diagnose HER2-Positive Breast Cancer

MOF derivatization of multifunctional nanozyme: Peroxidase-like catalytic activity combined with magnetic solid phase extraction for colorimetric detection of Hg2+

Nanozyme-based colorimetric sensing has drawn immense attention due to the rapid development of nanozyme in recent years. However, the selectivity of nanozyme-based colorimetric sensing greatly limits its subsequent practical application. It is well known that sample pretreatment can not only improve selectivity by eliminating the sample matrix interference, but also improve sensitivity by enriching trace targets. Based on the easy facile surface modification properties of nanozyme, we rationally designed nanozyme combined with sample pretreatment for colorimetric biosensing, through separation and enrichment, thereby improving the selectivity and sensitivity of the nanozyme colorimetric biosensing. As a proof of concept, the detection of Hg2+ by nanozyme-based colorimetric sensing was used as an example. Magnetic peroxidase-like nanozyme Fe3S4 was designed and synthesized. The selectivity is improved by the specific adsorption of S-Hg bond and the interference elimination after magnetic separation. In addition, the sensitivity is improved by magnetic solid-phase extraction enrichment. Our established colorimetric sensing based on Fe3S4 nanozyme integrated sample pretreatment with an enrichment factor of 100 and the limit of detection (LOD) is 26 nM. In addition, this strategy was successfully applied to detect Hg2+ in environmental water samples. Overall, the strategy showed good selectivity and sensitivity, providing a new practical method for the application of nanozyme-based biosensing in sample pretreatment.

A label-free colorimetric biosensor utilizing natural material for highly sensitive exosome detection

Exosomes, extracellular vesicles secreted by cells, play a crucial role in intercellular communication by transferring information from source cells to recipient cells. These vesicles carry important biomarkers, including nucleic acids and proteins, which provide valuable insights into the parent cells' status. As a result, exosomes have emerged as noninvasive indicators for the early diagnosis of cancer. Colorimetric biosensors have garnered significant attention due to their cost-effectiveness, simplicity, rapid response, and reproducibility. In this study, we employ sporopollenin microcapsules (SP), a natural biopolymer material derived from pollen, as a substrate for gold nanoparticles (AuNPs). By modifying the SP-Au complex with CD63 aptamers, we develop a label-free colorimetric biosensor for exosome detection. In the absence of exosomes, the SP-Au complex catalyzes the oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB), resulting in a color change from colorless to blue. However, the addition of exosomes inhibits the catalytic activity of the SP-Au complex due to coverage of exosomes on AuNPs. This colorimetric biosensor exhibits high sensitivity and selectivity for exosome detection, with a detection limit of 10 particles/μL and a wide linear range of 10 - 108 particles/μL. Additionally, the SP-Au biosensor demonstrates remarkable resistance to serum protein adsorption and excellent catalytic stability even in harsh environments, making it highly suitable for clinical diagnostics.

Silicene's intriguing nanozymatic activity: Improved colorimetric and electrochemically supported colorimetric biosensing

Silicene is a material that is largely studied via theoretical models. In the practical world, lengthy incubation times and poor sensitivity of colorimetric detection methods have limited their implementation. This paper transforms silicene's theoretical advantages into practical applications, presenting it as a tangible solution for significantly improving colorimetric biosensing. More specifically, this paper presents a pivotal breakthrough by not only documenting the remarkable nanozymatic activity of silicene in oxidizing 3,3′,5,5′-Tetramethylbenzidine (TMB) with hydrogen peroxide (H2O2), but also leveraging silicene’s inherent electronic properties to enhance the speed and precision of the peroxidase reaction, thereby improving its performance and advancing applicability. Two detection mechanisms were tested to observe the influence of silicene’s electronic scope; one in the absence and the other in the presence of an externally supplied electric potential. Both were successful in the detection of H2O2 and dopamine. The latter proved to be more favorable as it not only shortened the incubation time from 30 min to 1.5 min but also exhibited lower Limit of Detection (LOD) values. To demonstrate, for dopamine, in a concentration range of 0 – 10 μM, the LOD was 4.13 μM without the electric potential and 2.21 μM with the electric potential. Similarly, in a second concentration range, the LOD values were 37.81 μM and 34.94 μM, respectively. Kinetic profiles indicated the superior affinity silicene has for TMB and H2O2. A selectivity test also indicated that the method has excellent selectivity towards dopamine.

Colorimetric biosensing of triglyceride in waste water using lipase immobilized ZnO nanoparticle enriched chitosan nanoconjugate

We constructed a liquid-phase colorimetric biosensor to detect Triglycerides (TG) contamination in water using a Porcine Pancreatic Lipase Immobilized Zinc – chitosan nanocomposite matrix with Phenolsulfonphthalein as the chromophore. The nanoparticle-augmented matrix was characterized using an SEM—the change in the chromophore colour from red to yellow indicated vegetable oil hydrolysis and fatty acid release. The biosensor showed a LOQ between 25–500 µg/mL at an operating temperature between 25–45 °C, over a pH range of 4–9 within 5 minutes. The biosensor showed resilience to samples contaminated up to 50 times the BIS permissible limits for iron, copper and manganese. The sensor showed repeatability of nearly 100% with the LOD 3 µg/mL, sensitivity 47.1 U/µg/cm2 and retained up to 80% of its activity till the 10th day when stored at 10 °C. A fully adept version of the biosensor can be applied to detect TG in water and wastewater.

Rational design of nanozyme with integrated sample pretreatment for colorimetric biosensing

Nanozymes have been widely used in the field of biosensing owing to their high stability, low cost, adjustable catalytic activity, and convenient modification. However, achieving high selectivity and sensitivity simultaneously in nanozyme-based colorimetric sensing remains a major challenge. Nanozymes are nanomaterials with enzyme-simulating activity that are often used as solid-phase adsorbents for sample pretreatment. Our design strategy integrated sample pretreatment function into the nanozyme through separation and enrichment, thereby improving the selectivity and sensitivity of nanozyme-based colorimetric biosensing. As a proof-of-concept, glucose was used as the model analyte in this study. A phenylboric acid-modified magnetic nanozyme (Cu/Fe3O4@BA) was rationally designed and synthesized. Selectivity was enhanced by boronate-affinity specific adsorption and the elimination of interference after magnetic separation. In addition, magnetic solid-phase extraction enrichment was used to improve the sensitivity. A recovery rate of more than 80% was reached when the enrichment factor was 50. The synthesized magnetic Cu/Fe3O4@BA was recyclable at least five times. The proposed method exhibited excellent selectivity and sensitivity, simple operation, and recyclability, providing a novel and practical strategy for designing multifunctional nanozymes for biosensing.

DNA-functionalized cryogel based colorimetric biosensor for sensitive on-site detection of aflatoxin B1 in food samples

Hydrogel biosensors present numerous advantages in food safety analysis owing to their remarkable biocompatibility, cargo-loading capabilities and optical properties. However, the current drawbacks (slow target responsiveness and poor mechanical strength) restricted their further utilization at on-site detection of targets. To address these challenges, a DNA-functionalized cryogel with hierarchical pore structures is constructed to improve the reaction rate and the robustness of hydrogel biosensor. During cryogel preparation, ice crystals serve as templates, shaping interconnected hierarchical microporous structures to enhance mass transfer for faster responses. Meanwhile, in the non-freezing zone, concentrated monomers create a dense cross-linked network, strengthening cryogel matrix strength. Accordingly, a colorimetric biosensor based on DNA cryogel has been developed as a proof of concept for rapid detection of aflatoxin B1 (AFB1) in food samples, and an excellent analytical performance was obtained under the optimized conditions with a low detection limit (1 nM), broad detection range (5–100 nM), satisfactory accuracy and precision (recoveries, 81.2–112.6 %; CV, 2.75–5.53 %). Furthermore, by integrating with a smartphone sensing platform, a portable device was created for rapid on-site measurement of target within 45 min, which provided some insight for hydrogel biosensors design.

Smartphone-based device for rapid and single-step detection of piRNA–651 using anti-DNA:RNA hybrid antibody and enzymatic signal amplification

P-element-induced wimpy testis (PIWI)-interacting RNAs (piRNAs/piRs) are a class of small noncoding RNAs that play a crucial role in regulating various biological processes, including carcinogenesis. One specific piRNA, piR–651, has been reported to be overexpressed in both human blood serum and solid cancer tissues, that can be used a viable biomarker in cancer diagnosis. Early diagnosis of cancer can help reduce the burden of the disease and improve survival rates. In the present work, we report for the first time a smartphone-based colorimetric biosensor for highly sensitive and specific detection of piR–651 thanks to an enzymatic signal amplification, which yielded high colorimetric intensities. Indeed, a heteroduplex DNA:RNA was formed in the presence of piR–651 with the capture DNA probe immobilized on the magnetic beads for easy magnetic separation. Then, a HRP tethered to anti-DNA:RNA (S9.6) was used to reveal the DNA-RNA heteroduplex formed by catalyzing the oxidation of TMB substrate into colorimetric TMBox, which absorbs at 630 nm. The absorbance is positively proportional to the piR–651 concentrations. On the other hand, the colorimetric product of the assay can be photographed with a smartphone camera and analyzed using ImageJ software. Using a smartphone and under optimal conditions, the biosensor responded linearly to the logarithm of piRNA–651 from 8 fM to 100 pM with a detection limit of 2.3 fM and discriminates against other piRNAs. It was also successfully applied to the determination of piRNA–651 levels in spiked human serum.

A label free nano biosensor for the detection of E.coli using cysteine capped gold nanoparticle

This study presents a novel one-pot synthesis method to generate positively charged gold nanoparticles (+) Au NPs, utilizing the thiol-containing amino acid cysteine as a capping agent. The resulting Cys-Au NPs displayed a self-assembled monolayer of uniformly sized gold nanoparticles with the diameter of size 36 nM and demonstrated a distinct color change and absorbance shift from 530 to 512 nm when exposed to E. coli, enabling the development of a colorimetric biosensor for rapid and reliable detection of the bacteria. Cys-Au NPs were characterized using spectroscopy, SEM (Scanning electron microscopy); X-ray diffractometer analysis and the photophysical properties of the particles towards E. coli were investigated through absorption spectroscopy and colorimetric tests. The electrostatic binding between Cys-Au NPs and the negatively charged E. coli cell wall facilitated the sensor’s specificity. The biosensor exhibited a detection limit of 2.2 × 102 CFU/mL, with a linear detection range between 2.2 × 10¹ and 2.2 × 104 CFU/mL. The sensor’s applicability extends to the detection of E. coli in potable water, presenting significant potential in food safety and clinical diagnostics. This innovative detection method provides a quick and efficient means of identifying E. coli in water samples, offering broad applications in various fields, including rapid bacterial detection, food safety, and clinical diagnostics.

Synthesis of a COF-on-MOF hybrid nanomaterial for enhanced colorimetric biosensing

The construction of hybrid materials is significant for the exploration of functionalities in colorimetric biosensing due to its structural designability and synergy effects. In this work, a COF-on-MOF hybrid nanomaterial has been newly synthesized for colorimetric biosensing. Experimental results reveal that on-surface synthesis of COF on MOF brings nanoscale proximity between COF and MOF, which exhibits more than two folds of peroxidase-like activity as compared to single Fe-MOF. Therefore, by using the MCA@Fe-MOF nanomaterial with the assist of a specific acetyl-peptide, MCA@Fe-MOF can serve as an efficient signal reporter for colorimetric assay of histone deacetylase (HDAC), and the limit of detection (LOD) can be as low as 0.261 nM. Looking forward, the demand for diverse and promising COF-on-MOF nanomaterials with varied functionalities is anticipated, propelling further exploration of their role in colorimetric biosensing.

Improving the sensing sensitivity of silver nanoparticle-based colorimetric biosensors from the point of salt.

The sensing sensitivity was improved for silver nanoparticles (AgNPs)-based colorimetric biosensors by using the most suitable salt to induce AgNPs aggregation. As for the salt composed of low-affinity anion and monovalent cation, the cation-dependent charge screening effect was the driving force for AgNPs aggregation. Apart from the charge screening effect, both the bridging of multivalent cation to the surface ligand of AgNP and the interaction between anion and Ag contributed to inducing AgNPs aggregation. Considering the higher aggregation efficiency of AgNPs resulted in a narrower sensing range, salt composed of low-affinity anion and monovalent cation was recommended for AgNPs-based colorimetric analysis, which was confirmed by fourfold higher sensitivity of DNA-21 detection using NaF than NaCl. This work inspires further thinking on improving the sensing performanceof metal nanomaterials-based sensors from the point of colloidal surface science.

Trace Amount of Bi-Doped Core-Shell Pd@Pt Mesoporous Nanospheres with Specifically Enhanced Peroxidase-Like Activity Enable Sensitive and Accurate Detection of Acetylcholinesterase and Organophosphorus Nerve Agents.

The urgent need for sensitive and accurate assays to monitor acetylcholinesterase (AChE) activity and organophosphorus pesticides (OPs) arises from the imperative to safeguard human health and protect the ecosystem. Due to its cost-effectiveness, ease of operation, and rapid response, nanozyme-based colorimetry has been widely utilized in the determination of AChE activity and OPs. However, the rational design of nanozymes with high activity and specificity remains a great challenge. Herein, trace amount of Bi-doped core-shell Pd@Pt mesoporous nanospheres (Pd@PtBi2) have been successfully synthesized, exhibiting good peroxidase-like activity and specificity. With the incorporation of trace bismuth, there is a more than 4-fold enhancement in the peroxidase-like performance of Pd@PtBi2 compared to that of Pd@Pt. Besides, no significant improvement of oxidase-like and catalase-like activities of Pd@PtBi2 was found, which prevents interference from O2 and undesirable consumption of substrate H2O2. Based on the blocking impact of thiocholine, a colorimetric detection platform utilizing Pd@PtBi2 was constructed to monitor AChE activity with sensitivity and selectivity. Given the inhibition of OPs on AChE activity, a biosensor was further developed by integrating Pd@PtBi2 with AChE to detect OPs, capitalizing on the cascade amplification strategy. The OP biosensor achieved a detection limit as low as 0.06 ng mL-1, exhibiting high sensitivity and anti-interference ability. This work is promising for the construction of nanozymes with high activity and specificity, as well as the development of nanozyme-based colorimetric biosensors.

Optimizing single-atom cerium nanozyme activity to function in a sequential catalytic system for colorimetric biosensing

Ever-increasing attention is being drawn towards single-atom nanozymes (SAzymes) for colorimetric biosensing applications. A series of atomically dispersed cerium (Ce) nanozymes are synthesized with different feeding molar ratios of Ce/Zn or under various pyrolysis temperatures, and their enzymatic performance, including peroxidase-, oxidase-, and catalase-activities, is explored. Both theoretical and experimental results demonstrate that Ce species symmetrically coordinate with nitrogen atoms to form CeN4 moieties which exhibit peroxidase-like activity. The best Ce-based SAzyme achieves remarkable peroxidase-like specificity and enables the colorimetric sensing of H2O2. This sensitivity provides a versatile reporter platform for detecting the activities of natural enzymes involved in H2O2 generation and further monitoring their substrates as well as inhibitors, for example, organophosphorus pesticides (OPs) when integrating with the acetylcholine esterase-choline oxidase biocatalyzed cascade. More importantly, this proposed sequential catalytic biosensor exhibits the capability to monitor trace OPs in the real samples with high sensitivity yet insignificant interference. This work sheds new light on further design of highly active lanthanide-based SAzymes to boost enzyme-mimicking reactions for the applications in rapid on-site colorimetric analysis of emerging contaminants in environmental and food samples.

Colorimetric detection of circulating tumor cells in breast cancer based on ladder-branch hybridization chain reaction and DFs/AuNCs nanozyme

Breast cancer is the most common malignant tumor in women, which accounts for 6.9% of all cancer-related deaths. Early diagnosis is crucial for making the best clinical decision and improving the prognosis of patients. Circulating tumor cells (CTCs) have been regarded as significant tumor biomarkers. Herein, we designed a colorimetric biosensor for breast cancer CTCs quantification based on ladder-branch hybridization chain reaction (HCR) and DNA flowers/gold nanoclusters (DFs/AuNCs) nanozyme. With the assistance of complementary DNA labeled on magnetic beads (MBs), the cleavage products of RNA-cleaving DNAzymes (RCDs) could be rapidly captured, subsequently triggering ladder-branch HCR. In addition, the DFs/AuNCs nanozyme was applied for colorimetric analysis, which further improved the sensitivity for the detection of target CTCs. Benefiting from specific RCDs, ladder-branch HCR and DFs/AuNCs, we achieved a superior detection limit of 3 cells/mL as well as a broad linear range of 10 cells/mL to 104 cells/mL. Conclusively, this colorimetric biosensor achieved sensitively and selectively detection of breast cancer CTCs without the participation of enzymes at room temperature, which might provide new insight into the early detection of breast cancer.

Gold Nanoparticle-based Biosensors for Point-of-Care Diagnostics: A Review of Sensing Nanoparticle Applications and Future Prospects.

Point of Care Diagnostics (POCD) is quintessential in hospitals and the healthcare sector as the secants uplift the quality of medical care and the life of a patient by facilitating quick identification of the underlying pathological condition. Nanotechnology can provide opportunities and has potential in the development of new-age sensing/diagnostic tools. Owing to extraordinary features (e.g., higher density, effective catalysis, good conduction, biocompatibility, inertness, and greater surface-to-volume ratio), gold nanoparticles (AuNPs) are frequently employed in POCT (Point-of-Care-Testing). Gold nanoparticles-based colorimetric methods are widely used in the rapid, sensitive, and selective detection of analytes/target molecules. AuNPs description is critical for their possible utility in prophylaxis, diagnostics, and treatment of an ailment. AuNPs interact with organic/inorganic target molecules to generate colorimetric shift that enables the accurate, precise, and subtle recognition of biologicals (e.g., microorganisms, cellular components, and proteins) and metal ions. This review focused on the need for AuNPs-based colorimetric application in prophylaxis, diagnostics, and treatment in healthcare and reviewed the future outlook of these AuNPs for biological applications. Different synthesis methods of AuNPs, their morphology, and characterization, including their surface functionalization, will be discussed in detail. AuNPs are very much preferable nanomaterials owing to exclusive optical, electrical, and photothermal features. AuNPs-based colorimetric biosensors are simple and possess great utility, yet these offer a robust technique to enable visual, quantitative analysis.

Engineered nanozymes for speedy colorimetric biosensing and sensitive cascaded-catalytic determining of norfloxacin

Nanozymes based on default engineering have garnered considerable interest for their ability to effectively imitate natural enzyme catalysis settings for detection. With the widespread use of quinolone antibiotics, it is worth noting that the antibiotic drugs represented by norfloxacin (NOR) are concentration-dependent when used. Therefore, residual or dosage differences in the body can lead to some central toxicity hazards, so monitoring and detection are necessary. Herein, we reported Fe3O4@MIL-101@polypyrroles (PPys) nanozymes with default engineering, which possessed a porous core–shell structure with Fe3O4 as the core and MIL-101 and PPys as the shells in sequence. X-Ray photoelectron spectroscopy (XPS) analysis and density functional theory (DFT) calculation were to guide and analyze the catalytic pathway of peroxidase-like activity. Furthermore, a versatile intelligent colorimetric and sensing platform has been established to achieve sensitive cascaded-catalytic determining norfloxacin (NOR). In addition, NOR has a lower threshold of acceptability detection of 0.14 μM, which is better than the interpreted detection methods. We have firstly explored the detection mechanisms of ascorbate (H2A) and NOR and discovered that NOR could selectively oxidize Fe2+ and support Bi-Bi mechanisms in the process of H2O2 catalysis. Excellent guidelines for the design of multifunctional nanozyme catalysts were provided by the inquiry.

Colorimetric biosensor based on aptamer recognition-induced multi-DNA release and peroxidase-mimicking three-way junction DNA-Ag/PtNCs for the detection of Salmonella typhimurium

Salmonella typhimurium, as a major foodborne pathogen, poses a serious threat to public health safety worldwide. Here, we present a colorimetric biosensor based on aptamer recognition-induced multi-DNA release and peroxidase-mimicking three-way junction DNA-silver/platinum bimetallic nanoclusters (3WJ/DNA-Ag/PtNCs) for the detection of S. typhimurium. In this method, S. typhimurium specifically binds to the aptamer and releases multiple cDNAs to form the three-way junction DNA structure and synthesize silver/platinum bimetallic nanoclusters, which induces signaling changes. Interestingly and importantly, the use of 3WJ/DNA as the template for synthesizing Ag/PtNCs gives the method an extremely low background signal. Under the optimal conditions, the constructed biosensor had a linear response range of 2.6 × 102-2.6 × 106 CFU/mL and a detection limit of 2.6 × 102 CFU/mL for the detection of S. typhimurium. In addition, the proposed method can effectively detect S. typhimurium in milk.

Ultrasensitive Nanofiber Biosensor: Rapid In Situ Chromatic Detection of Bacteria for Healthcare Innovation.

Rapid detection of bacterial presence in skin wounds is crucial to prevent the transition from acute to chronic wounds and the onset of systemic infections. Current methods for detecting infections, particularly at low concentrations (<1.0 × 105 CFU/cm2), often require complex technologies and direct sampling, which can be invasive and time-consuming. Addressing this gap, we introduce a colorimetric nanofibrous biosensor enabling real-time in situ monitoring of bacterial concentrations in wounds. This biosensor employs a colorimetric hemicyanine dye (HCy) probe, which changes color in response to bacterial lipase, a common secretion in infected wounds. To enhance the biosensor's sensitivity, we incorporated two key materials science strategies: aligning the nanofibers to promote efficient bacterial attachment and localization and integrating Tween 80, a surfactant, within the nanofiber matrix. This combination of physical and chemical cues results in a notable increase in lipase activity. The cross-aligned core-shell nanofibers, embedded with Tween 80 and HCy, demonstrate an immediate and distinct color change when exposed to as low as 3.0 × 104 CFU/cm2 of common pathogens such as Pseudomonas aeruginosa and MRSA. Significantly, the presence of Tween 80 amplifies the colorimetric response, making visual detection more straightforward and four times more pronounced. Our nanobiosensor design facilitates the detection of low-concentration bacterial infections in situ without the need to remove wound dressings. This advancement marks a significant step forward in real-time wound monitoring, offering a practical tool for the early detection of clinical bacterial infections.

Sensitive colorimetric aptasensor for multiple foodborne pathogens detection based on PCN-Mo peroxidase-like activity

A rapid and sensitive colorimetric biosensor was developed for simultaneous detection of various pathogenic bacteria based on aptamer-mediated peroxidase (POD)-like activity of PCN-Mo nanozyme. Initially, a porphyrin-based PCN-Mo metal organic framework (MOF) nanozyme is constructed through a simple hydrothermal method, as thoroughly figured out by a series of characterization techniques. The study showed that the pathogenic bacteria aptamers could adhere to the surface of PCN-Mo, and significantly enhanced the POD-like activity by reinforcing their transport of electrons to 3,3′,5,5′-tetramethylbenzidine (TMB). In the presence of different target bacteria, the aptamers tend to bind to bacteria specifically, causing the further enhancement of POD-like activity. Based on the signal-amplification by the PCN-Mo@aptamer (PCN-Mo@Apt) nanozyme, the constructed pathogenic bacteria sensor demonstrated a wide linear range between 101 to 105 CFU/mL, with the detection limits (LODs) of 0.578, 0.439, and 0.643 CFU/mL for E. coli, S. typhimurium and S. aureus, respectively. Furthermore, the PCN-Mo sensor was capable of detection pathogenic bacteria in water, milk, classic milk tea, chicken, and beef with the recoveries of 90.1%–104.1%, indicating the sensor's practicability for pathogenic bacteria detection in real samples.