Rapid Detection Of Pathogenic Bacteria Research Articles

Femtosecond laser-induced fluorescence spectroscopy for the rapid detection of pathogenic bacteria

Femtosecond laser-induced fluorescence spectroscopy for the rapid detection of pathogenic bacteria

Rapid detection of nucleic acid sequences of pathogenic bacteria based on a series piezoelectric quartz crystal sensor with transcription activator-like effectors

Accurate and rapid detection of pathogenic bacteria is of great importance in the field of clinical diagnosis and food safety. Current methods for pathogenic bacteria detection have some problems in accurate, rapid and universal application. Here we proposed a pathogenic bacteria series piezoelectric quartz crystal (SPQC) sensor for achieving highly specific and sensitive detection of pathogenic bacteria. The universal sequences of common clinical pathogens screened by our group were used as detection targets. A new TALEs nuclease was synthesized as a recognition element, which recognizes double-stranded DNA at the level of a single base mismatch in the range of 17–19 bases. Targets could be specifically recognized by TALEs, resulting in the change of electrode surface, which would be further amplified by hybridization chain reaction and silver staining technique. Finally, the changes would be detected by SPQC system. This strategy was demonstrated to have excellent performance, enabling sensitive detection of targets with a detection limit of 25 cfu/mL in less than 3 h. What's more, the identification of single base mismatch could be achieved when the target ranging in length between 17 and 19 bases. The proposed method is rapid, accurate and easy universal application and expected to be applied in clinical diagnosis and food safety.

A Novel Dual Bacteria-Imprinted Polymer Sensor for Highly Selective and Rapid Detection of Pathogenic Bacteria.

The rapid, sensitive, and selective detection of pathogenic bacteria is of utmost importance in ensuring food safety and preventing the spread of infectious diseases. Here, we present a novel, reusable, and cost-effective impedimetric sensor based on a dual bacteria-imprinted polymer (DBIP) for the specific detection of Escherichia coli O157:H7 and Staphylococcus aureus. The DBIP sensor stands out with its remarkably short fabrication time of just 20 min, achieved through the efficient electro-polymerization of o-phenylenediamine monomer in the presence of dual bacterial templates, followed by in-situ template removal. The key structural feature of the DBIP sensor lies in the cavity-free imprinting sites, indicative of a thin layer of bacterial surface imprinting. This facilitates rapid rebinding of the target bacteria within a mere 15 min, while the sensing interface regenerates in just 10 min, enhancing the sensor's overall efficiency. A notable advantage of the DBIP sensor is its exceptional selectivity, capable of distinguishing the target bacteria from closely related bacterial strains, including different serotypes. Moreover, the sensor exhibits high sensitivity, showcasing a low detection limit of approximately 9 CFU mL-1. The sensor's reusability further enhances its cost-effectiveness, reducing the need for frequent sensor replacements. The practicality of the DBIP sensor was demonstrated in the analysis of real apple juice samples, yielding good recoveries. The integration of quick fabrication, high selectivity, rapid response, sensitivity, and reusability makes the DBIP sensor a promising solution for monitoring pathogenic bacteria, playing a crucial role in ensuring food safety and safeguarding public health.

Dual Synthetic Receptor-Based Sandwich Electrochemical Sensor for Highly Selective and Ultrasensitive Detection of Pathogenic Bacteria at the Single-Cell Level.

Sensitive and rapid detection of pathogenic bacteria is essential for effective source control and prevention of microbial infectious diseases. However, it remains a substantial challenge to rapidly detect bacteria at the single-cell level. Herein, we present an electrochemical sandwich sensor for highly selective and ultrasensitive detection of a single bacterial cell based on dual recognition by the bacteria-imprinted polymer film (BIF) and aptamer. The BIF was used as the capture probe, which was in situ fabricated on the electrode surface within 15 min via electropolymerization. The aptamer and electroactive 6-(Ferrocenyl)hexanethiol cofunctionalized gold nanoparticles (Au@Fc-Apt) were employed as the signal probe. Once the target bacteria were anchored on the BIF-modified electrode, the Au@Fc-Apt was further specifically bound to the bacteria, generating enhanced current signals for ultrasensitive detection of Staphylococcus aureus down to a single cell in phosphate buffer solution. Even in the complex milk samples, the sensor could detect as low as 10 CFU mL-1 of S. aureus without any complicated pretreatment except for 10-fold dilution. Moreover, the current response to the target bacteria was hardly affected by the coexisting multiple interfering bacteria, whose number is 30 times higher than the target, demonstrating the excellent selectivity of the sensor. Compared with most reported sandwich-type electrochemical sensors, this assay is more sensitive and more rapid, requiring less time (1.5 h) for the sensing interface construction. By virtue of its sensitivity, rapidity, selectivity, and cost-effectiveness, the sensor can serve as a universal detection platform for monitoring pathogenic bacteria in fields of food/public safety.

Cutting-Edge Advancements in EIS Technologies for Rapid Detection of Pathogenic Bacteria in Water

The emerging research surrounding biosensors has seen an unprecedented rise in recent years: from innovative methods of exploiting the bio-recognition event to groundbreaking first-generation designs, work around biosensors is playing a more substantial role in all facets of modern biotechnology. This mini-review explores a type of biosensor that analyzes bio-recognition events through a technique known as electrochemical impedance spectroscopy (EIS). The EIS technique belongs to the electrochemical class of biosensors and is used to examine analyte-electrode interactions through the transfer of electrons. While the technique has proved effective in detecting several virulent bacteria, this review will primarily focus on E. coli O157:H7, L.monocytogenes, and S. Typhimurium. These three pathogens are all highly contagious and capable of causing severe infections and thus must be carefully managed in essential resources such as water. Though existing methods are effective, there are ways EIS biosensing can be further enhanced in terms of accuracy and precision. At the end of the manuscript, we further overview the state-of-the-art challenges and opportunities in EIS.

Rapid detection of pathogenic bacteria based on a universal dual-recognition FRET sensing system constructed with aptamer-quantum dots and lectin-gold nanoparticles

The threat to public health from bacterial infections has led to an urgent need to develop simpler, faster and more reliable bacterial detection methods. In this work, we developed a universal dual-recognition based sandwich fluorescence resonance energy transfer (FRET) sensor by using specific aptamer-modified quantum dots (Aptamer-QDs) as energy donor and lectin concanavalin A (Con A) modified gold nanoparticles (Con A-AuNPs) as energy acceptor to achieve rapid and sensitive detection of Escherichia coli (E. coli) within 0.5 h. In the presence of the target E. coli, the energy donor of Aptamer-QDs and acceptor of Con A-AuNPs were close to each other, causing changes of FRET signals. Based on the constructed FRET sensor, a linear detection range of from 102 cfu/mL to 2 × 108 cfu/mL with the detection limit of 45 cfu/mL for E. coli was achieved. Furthermore, the FRET sensor was applied to detect E. coli in the milk and orange juice with the detection limit of 300 cfu/mL and 200 cfu/mL, respectively and recovery rate from 83.1% to 112.5%. The strategy holds great promise in pathogenic bacteria detection due to its rapid and sensitivity.

Optical Methods for Label-Free Detection of Bacteria

Pathogenic bacteria are the leading causes of food-borne and water-borne infections, and one of the most serious public threats. Traditional bacterial detection techniques, including plate culture, polymerase chain reaction, and enzyme-linked immunosorbent assay are time-consuming, while hindering precise therapy initiation. Thus, rapid detection of bacteria is of vital clinical importance in reducing the misuse of antibiotics. Among the most recently developed methods, the label-free optical approach is one of the most promising methods that is able to address this challenge due to its rapidity, simplicity, and relatively low-cost. This paper reviews optical methods such as surface-enhanced Raman scattering spectroscopy, surface plasmon resonance, and dark-field microscopic imaging techniques for the rapid detection of pathogenic bacteria in a label-free manner. The advantages and disadvantages of these label-free technologies for bacterial detection are summarized in order to promote their application for rapid bacterial detection in source-limited environments and for drug resistance assessments.

Label-free single-particle imaging approach for ultra-rapid detection of pathogenic bacteria in clinical samples

Rapid detection of pathogenic bacteria within a few minutes is the key to control infectious disease. However, rapid detection of pathogenic bacteria in clinical samples is quite a challenging task due to the complex matrix, as well as the low abundance of bacteria in real samples. Herein, we employ a label-free single-particle imaging approach to address this challenge. By tracking the scattering intensity variation of single particles in free solution, the morphological heterogeneity can be well identified with particle size smaller than the diffraction limit, facilitating the morphological identification of single bacteria from a complex matrix in a label-free manner. Furthermore, the manipulation of convection in free solution enables the rapid screening of low-abundance bacteria in a small field of view, which significantly improves the sensitivity of single-particle detection. As a proof of concept demonstration, we are able to differentiate the group B streptococci (GBS)-positive samples within 10 min from vaginal swabs without using any biological reagents. This is the most rapid and low-cost method to the best of our knowledge. We believe that such a single-particle imaging approach will find wider applications in clinical diagnosis and disease control due to its high sensitivity, rapidity, simplicity, and low cost.

Application of nucleic acid aptamer-based lateral flow assay in microbe detection

<p indent="0mm">Microbial contamination and infection are global issues in the food and environmental fields that seriously threaten human health. Bacteria and fungi can easily cause food spoilage, resulting in diarrhea and vomiting; viruses can infect humans through different transmission routes, causing severe or even fatal harm. Hence, rapid analysis and identification of pathogenic microorganisms and simultaneous detection of multiple types of microbes have become hot research topics in biochemical analysis and molecular diagnosis. The lateral flow assay (LFA) is a simple, rapid, economical, and efficient detection technology with high sensitivity, simple operation, and environmental friendliness. It can provide instant test results under non-laboratory circumstances, hence becoming an ideal choice for point-of-care testing, which has been applied to rapidly detect various targets. The current conventional principle of the LFA is still based on the specific recognition of the antigen by the antibody. However, as a commonly used target recognition molecule in conventional biochemical and medical detection, the application of antibodies also has certain limitations for rapid and accurate identification of certain targets due to strict control of the production and purification process, as well as susceptibility to the interference of the operating environment, pH, temperature, and other conditions, such as long production cycle, high cost, poor stability, and cross-reactivity. Aptamers are a class of single-stranded DNA (ssDNA) or RNA obtained through the systematic evolution of ligands by exponential enrichment (SELEX), which can usually form a stable secondary structure. Aptamers can be folded into a three-dimensional structure through conformational change and interact with the target through conformation complementarity, π-π stacking between aromatic rings, base stacking, electrostatic interaction, and hydrogen bonding. So far, nearly 300 kinds of aptamers have been discovered. As alternatives, aptamers are easy and facile to modify and label with high sensitivity and specificity. Accordingly, the innovative rapid detection technique can be developed by combining the LFA with an aptamer. This aptamer-based LFA technology can be widely used in qualitative, semi-quantitative, and quantitative detection in food safety, environment, clinical, and other fields. Nowadays, most microbe detection methods are constructed based on this approach. The common strategies of aptamer-based LFAs include the sandwich method, competitive method, and adsorption–desorption method. Diverse ingenious materials such as gold nanoparticles and quantum dots have also been proposed for signal read-out. Different signal capture models, such as colorimetric and fluorescence methods, have been applied for sensitive and accurate detection of a single or multiple target microbe. Furthermore, in view of the unique properties of nucleic acid aptamers, several signal amplification methods can be further involved in the LFA to enhance the sensitivity for target detection. This review introduces the use of aptamers with different structural patterns and labeling types in recent years, as well as a variety of methods to detect microbes, especially for the rapid detection of pathogenic bacteria. Based on the excellent characteristics, the aptamer-based LFA presents more flexibility and selectivity for microbe detection with good applicability, specificity, and sensitivity and can better achieve low-cost, rapid detection. This study is expected to provide a reference for developing nucleic acid aptamer-based LFA technologies, especially for efficient and accurate diagnosis of corona virus disease 2019 (COVID-19), exploiting the novel application scope of LFA technology.

Smartphone readable colorimetry and ICP-MS dual-mode sensing platform for ultrasensitive and label-free detection of Escherichia coli based on filter-assisted separation

The development and application of portable and user-friendly biosensing technology for rapid detection of pathogenic bacteria are essential for human and environmental care. In this work, based on the phenomenon that the filter membrane can selectively separate MnO2 nanosheets (MnO2 NSs) and Mn2+, a novel label-free sensing platform was developed to detect the Escherichia coli (E. coli) using β-galactosidase (β-Gal) as a marker. MnO2 NSs were utilized as a dual-mode signal molecule of smartphone-based colorimetric analysis and inductively coupled plasma mass spectrometry (ICP-MS). β-Gal can catalyze the 4-Aminophenyl β-D-galactopyranoside (PAPG) to produce p-aminophenol (PAP), which could reduce MnO2 NSs to Mn2+. After selective separation of MnO2 NSs and Mn2+ by the filter, colorimetric detection was achieved with a smartphone by identifying the RGB value of the MnO2 NSs coated membrane, while the Mn2+ in the filtrate was detected by ICP-MS. Under optimal conditions, the limits of detection (LODs) of E. coli by smartphone-based RGB analysis and ICP-MS were 5.6 × 103 CFU mL−1 and 35 CFU mL−1, respectively. This method was successfully used for E. coli assay in meat, vegetables, and fruit samples with the advantages of sensitivity, simplicity, and short incubation time (within 1 h).

In Situ Collection and Rapid Detection of Pathogenic Bacteria Using a Flexible SERS Platform Combined with a Portable Raman Spectrometer.

This study aims to develop a simple, sensitive, low-cost, environmentally friendly and flexible surface-enhanced Raman scattering (SERS) platform, combined with a portable Raman spectrometer, for the rapid and on-site SERS detection of bacteria. Commercial tobacco packaging paper (TPP) with little background interference was used as a loading medium that effectively adsorbed Au nanoparticles and provided sufficient “hot spots”. This Au-tobacco packaging paper (Au-TPP) substrate used as a flexible SERS platform can maximize sample collection by wiping irregular surfaces, and was successfully applied to the on-site and rapid detection of pathogenic bacteria. Raman fingerprints of pathogenic bacteria can be obtained by SERS detection of spiked pork using wipeable Au-TPP, which verifies its value in practical applications. The results collected by SERS were further verified by polymerase chain reaction (PCR) results. It showed several advantages in on-site SERS detection, including accurate discrimination, simple preparation, easy operation, good sensitivity, accuracy and reproducibility. This study indicates that the established flexible SERS platform has good practical applications in pathogenic bacterial identification and other rapid detections.

Immunofluorescent-aggregation assay based on anti-Salmonella typhimurium IgG-AuNCs, for rapid detection of Salmonella typhimurium.

Sensitive and rapid detection of pathogenic bacteria plays an important role in avoiding food poisoning. However, the practical application value of conventional assays for detection of foodborne bacteria, are limited by major drawbacks; these include the laboriousness of pure culture preparation, complexity of DNA extraction for polymerase chain reaction, and low sensitivity of enzyme-linked immunosorbent assay. Herein, we designed a non-complex strategy for the sensitive, quantitative, and rapid detection of Salmonella typhimurium with high specificity, using an anti-Salmonella typhimurium IgG-AuNC-based immunofluorescent-aggregation assay. Salmonella typhimurium was agglutinated with fluorescent anti-Salmonella typhimurium IgG-AuNC on a glass slide, and observed using a fluorescence microscope with photoexcitation and photoemission at 560nm and 620nm, respectively. Under optimized reaction conditions, the AuNC-based immunofluorescent-aggregation assay had a determinationrange between 7.0 × 103 and 3.0 × 108CFU/mL, a limit of detection of 1.0 × 103CFU/mL and an assay response time of 3min. Thetechnique delivered good results in assessing real samples.

Metagenomic Next-Generation Sequencing for the Microbiological Diagnosis of Abdominal Sepsis Patients.

ObjectivesFor patients with intra-abdominal infection (IAI), the rapid and accurate identification of pathogens remains a challenge. Metagenomic next-generation sequencing (mNGS) is a novel technique for infectious diseases, but its application in IAI is limited. In this study, we compared the microbiological diagnostic ability of plasma mNGS with that of conventional peritoneal drainage (PD) culture in critical care settings.MethodsFrom January 2018 to December 2020, a prospective observational study was performed at a tertiary teaching hospital in China and data on 109 abdominal sepsis patients were collected. The pathogen detection performance of plasma mNGS and PD culture method were compared.Measurements and ResultsNinety-two positive cases detected on PD culture, while plasma mNGS detected 61 positive cases. Forty-five patients (44.0%) had at least one matched pair of plasma mNGS and PD culture results. Compared with PD culture, the plasma mNGS was more rapid (27.1 ± 4.0 vs. 68.9 ± 22.3 h, p < 0.05). The patients received initial antibiotic treatment matched with mNGS detection showed better clinical outcomes.ConclusionFor abdominal sepsis patients, plasma mNGS can provide early, noninvasive, and rapid microbiological diagnosis. Compared with conventional PD smear, culture, and blood culture methods, plasma mNGS promote the rapid detection of pathogenic bacteria.

Reverse Transcription Loop-Mediated Isothermal Amplification Assay with High Sensitivity to Rapid Detection of Viable Salmonella in Foods

Background: Salmonella is one of the main foodborne bacterial pathogens, causing diseases and death. The study used reverse transcription loop-mediated isothermal amplification (RT-LAMP) to detect Salmonella. Objectives: To design six primers and detect Salmonella using RT-LAMP to facilitate the rapid detection of pathogenic bacteria in food. Methods: We designed six primers based on the gene coding sequences of inv A, specific to Salmonella. Each reaction solution contained 6.0 mM MgSO4, 1 M betaine, 1.6 mM dNTPs, 160 U/mL Bst DNA polymerase, 0.2 μM of both external primers, 0.8 μM of both internal primers, and 0.2 μM of both loop primers. The reaction temperature was 65°C. Results: Our amplified products were separated by 2% agarose gel electrophoresis. The detection limit was 10 CFU per reaction. Conclusions: RT-LAMP exhibited the same accuracy as the Chinese National Standard assay (GB 4789.30-2016) assay in detecting Salmonella in foods. RT-LAMP was highly specific and sensitive; hence, it may serve as an effective tool in detecting Salmonella.

A high-efficiency pretreatment method for elution of pathogenic bacteria in lettuce

Abstract Many current studies on rapid detection of pathogenic bacteria in foods have focused on the construction of detection methods, neglecting pretreatment. It is also a key step to efficiently elute pathogenic bacteria from food samples for rapid detection and can even determine the success or failure of an assay. In this study, we used Escherichia coli (E. coli), Salmonella enteritidis (S. enteritidis), and Listeria monocytogenes (L. monocytogenes) as model bacteria to compare the elution efficiency of different eluants; explore the effect of surfactant, ionic strength, protein (or amino acid and peptide), and enzyme on the recovery rate of bacteria in lettuce; and compare the compound effect caused by multiple factors. Finally, we developed an efficient bacterial recovery method and confirmed the superiority of this method to analyze the bacterial diversity of eluants from lettuce. The results showed that the recovery efficiency of E. coli, S. enteritidis, and L. monocytogenes, which were artificially contaminated with approximately 105 CFU/g in lettuces, could reach 94.4%, 90.6%, and 93.7% by using 10 mmol/L Tris·HCl (pH 9.5) with 0.1% peptone and 300 U/100 mL of cellulase, and furthermore, the elution efficiency could reach 99.6%, 98.6%, and 100% with the aid of a 2-min stomaching. For the lettuce samples with only native bacteria, the recovery rate reached 92.1% for viable aerobic bacteria by this method, which was approximately 10% higher than that of the modified previous method. The bacterial diversity of the eluted solution was analyzed, and the result was significantly improved. Considering these advantages, it is important to improve the elution efficiency to achieve rapid and accurate detection of pathogenic bacteria in lettuces.

Rapid Detection of Pathogenic Bacteria by the Naked Eye.

Escherichia coli O157:H7 and Staphylococcus aureus are common pathogens. Gram-negative bacteria, such as E. coli, contain high concentrations of endogenous peroxidases, whereas Gram-positive bacteria, such as S. aureus, possess abundant endogenous catalases. Colorless 3,5,3′,5′-tetramethyl benzidine (TMB) changes to blue oxidized TMB in the presence of E. coli and a low concentration of H2O2 (e.g., ~11 mM) at pH of 3. Moreover, visible air bubbles containing oxygen are generated after S. aureus reacts with H2O2 at a high concentration (e.g., 180 mM) at pH of 3. A novel method for rapidly detecting the presence of bacteria on the surfaces of samples, on the basis of these two endogenous enzymatic reactions, was explored. Briefly, a cotton swab was used for collecting bacteria from the surfaces of samples, such as tomatoes and door handles, then two-step endogenous enzymatic reactions were carried out. In the first step, a cotton swab containing bacteria was immersed in a reagent comprising H2O2 (11.2 mM) and TMB for 25 min. In the second step, the swab was dipped further in H2O2 (180 mM) at pH 3 for 5 min. Results showed that the presence of Gram-negative bacteria, such as E. coli with a cell number of ≥ ~105, and Gram-positive bacteria, such as S. aureus with a cell number of ≥ ~106, can be visually confirmed according to the appearance of the blue color in the swab and the formation of air bubbles in the reagent solution, respectively, within ~30 min. To improve visual sensitivity, we dipped the swab carrying the bacteria in a vial containing a growth broth, incubated it for ~4 h, and carried out the two-stage reaction steps. Results showed that bluish swabs resulting from the presence of E. coli O157: H7 with initial cell numbers of ≥ ~34 were obtained, whereas air bubbles were visible in the samples containing S. aureus with initial cell numbers of ≥ ~8.5 × 103.

Recent progress on microfluidic biosensors for rapid detection of pathogenic bacteria

Rapid on-site detection of pathogenic bacteria with high sensitivity and specificity is becoming an urgent need in public health assurance, medical diagnostics, environmental monitoring, and food safety fields. Despite being reliable and widely used, the existing methods of bacteria detection are cumbersome and time-consuming, which is not conducive to field detection. Microfluidic lab-on-a-chip technology has provided a detective tool for various analytes, due to its miniaturization, portability and low reagent consumption. Within this progress report, advances in the bacteria detection using microfluidic biosensors were discussed. Typical methods for pathogenic bacteria capture, separation and detection were introduced respectively in the first part. Then key applications of microfluidic biosensor-based rapid bacteria detection were presented. Finally, we made a conclusion and discussed possible research prospects in aspects of microfluidic biosensors for rapid detection of pathogenic bacteria.

Tailor-made magnetic nanocomposite with pH and thermo-dual responsive copolymer brush for bacterial separation

Rapid detection of pathogenic bacteria particularly in food samples demands efficient separation and enrichment strategies. Here, hydrophilic temperature-responsive boronate affinity magnetic nanocomposites were established for selective enrichment of bacteria. The thermo-responsive polymer brushes were developed by surface-initiated atom transfer radical polymerization of N-isopropylacrylamide (NIPAm) and allyl glycidyl ether (AGE), followed by a reaction of epoxy groups, and incorporation of fluorophenylboronic acid. The physical and chemical characteristics of the magnetic nanocomposites were analyzed systematically. After optimization, S. aureus and Salmonella spp. showed high binding capacities of 32.14 × 106 CFU/mg and 50.98 × 106 CFU/mg in 0.01 M PBS (pH 7.4) without bacteria death. Bacterial bindings can be controlled by altering temperature and the application of competing monosaccharides. The nanocomposite was then utilized to enrich S. aureus and Salmonella spp. from the spiked tap water, 25% milk, and turbot extraction samples followed by multiplex polymerase chain reaction (mPCR), which resulted in high bacteria enrichment, and demonstrated great potential in separation of bacteria from food samples.

Molecular imprinting technology for sensing foodborne pathogenic bacteria.

Foodborne diseases caused by bacterial pathogens pose a widespread and growing threat to public health in the world. Rapid detection of pathogenic bacteria is of great importance to prevent foodborne diseases and ensure food safety. However, traditional detection methods are time-consuming, labour intensive and expensive. In recent years, many attempts have been made to develop alternative methods for bacterial detection. Biosensors integrated with molecular imprinted polymers (MIPs) and various transducer platforms are among the most promising candidates for the detection of pathogenic bacteria in a highly sensitive, selective and ultra-rapid manner. In this review, we summarize the most recent advances in molecular imprinting for bacterial detection, introduce the underlying recognition mechanisms and highlight the applications of MIP-based biosensors. In addition, the challenges and future perspectives are discussed with the aim of accelerating the development of MIP-based biosensors and extending their applications.

3D Hierarchical Nanotopography for On-Site Rapid Capture and Sensitive Detection of Infectious Microbial Pathogens.

Effective capture and rapid detection of pathogenic bacteria causing pandemic/epidemic diseases is an important task for global surveillance and prevention of human health threats. Here, we present an advanced approach for the on-site capture and detection of pathogenic bacteria through the combination of hierarchical nanostructures and a nuclease-responsive DNA probe. The specially designed hierarchical nanocilia and network structures on the pillar arrays, termed 3D bacterial capturing nanotopographical trap, exhibit excellent mechanical reliability and rapid (<30 s) and irreversible bacterial capturability. Moreover, the nuclease-responsive DNA probe enables the highly sensitive and extremely fast (<1 min) detection of bacteria. The bacterial capturing nanotopographical trap (b-CNT) facilitates the on-site capture and detection of notorious infectious pathogens (Escherichia coli O157:H7, Salmonella enteritidis, Staphylococcus aureus, and Bacillus cereus) from kitchen tools and food samples. Accordingly, the usefulness of the b-CNT is confirmed as a simple, fast, sensitive, portable, and robust on-site capture and detection tool for point-of-care testing.