Search Results (55)

Foodborne pathogenic bacteria critically threaten public health and food industry sustainability, serving as a predominant trigger of food contamination incidents. To mitigate these risks, the development of rapid, sensitive, and highly specific detection technologies is essential for early warning and effective control of foodborne diseases. In recent years, biosensors have gained prominence as a cutting-edge tool for detecting foodborne pathogens, owing to their operational simplicity, rapid response, high sensitivity, and suitability for on-site applications. This review provides a comprehensive evaluation of critical biorecognition elements, such as antibodies, aptamers, nucleic acids, enzymes, cell receptors, molecularly imprinted polymers (MIPs), and bacteriophages. We highlight their design strategies, recent advancements, and pivotal contributions to improving detection specificity and sensitivity. Additionally, we systematically examine mainstream biosensor-based detection technologies, with a focus on three dominant types: electrochemical biosensors, optical biosensors, and piezoelectric biosensors. For each category, we analyze its fundamental principles, structural features, and practical applications in food safety monitoring. Finally, this review identifies future research priorities, including multiplex target detection, enhanced processing of complex samples, commercialization, and scalable deployment of biosensors. These advancements are expected to bridge the gap between laboratory research and real-world food safety surveillance, fostering more robust and practical solutions. Full article

Shaking off the forgetfulness towards the methodological power of inosine-mediated error-prone PCR (epPCR), this study reintroduces an often-underappreciated method as a considerably powerful approach for generating aptamer libraries from a single decameric ATCG-repeat-oligonucleotide. The aim was to demonstrate that this simple way of creating sequence diversity was suitable for delivering functional starting libraries for a set of ten whole-cell-SELEX (Systematic Evolution of Ligands by Exponential Enrichment) processes. This epPCR method uses inosine to introduce targeted mutations, avoiding the need for commercial oligo pools or large-scale synthesis. We applied this method to a “universal aptamer” and subjected the three resulting libraries to two rounds of selection against ten diverse targets including probiotic and pathogenic bacteria (Gram-negative and -positive) as well as human cell lines. The enriched aptamers exhibited new binding specificities, demonstrating that the approach supports functional selection. Much like dusting off an old tool and finding it perfectly suited for a modern task, this work shows that revisiting established techniques can address current challenges in aptamer development. Our main finding is that epPCR provides a robust, cost-effective strategy for generating starting libraries and lowers the barrier for initiating successful SELEX campaigns. Full article

Pseudomonas aeruginosa is a significant opportunistic pathogen highly prevalent in the environment, requiring early detection methods to prevent infections in vulnerable individuals. The most specific aptamer for P. aeruginosa, F23, has been used for the development of various assays and sensors for early diagnosis and monitoring. In this study, a novel F23-based electrochemical aptasensor was designed using disposal gold screen-printed electrodes (Au-SPEs) with high reproducibility. Methylene blue (MB) was used as an exogenous indicator, which significantly amplified the electrochemical signal and improved the sensitivity of detection. The aptasensor explored a limit of detection (LOD) of 8 CFU·mL⁻¹ and high selectivity for P. aeruginosa over other interfering bacteria. Furthermore, it showed potential to detect P. aeruginosa in tap water samples, offering a point-of-care tool for rapidly controlling the growth of this bacterium in various applications. Full article

Rapid and specific diagnosis is necessary for both the treatment and prevention of infectious diseases. Bacteria and viruses that enter the bloodstream can trigger a strong immune response in infected animals and humans. The fluorescence polarization assay (FPA) is a rapid and accurate method for detecting specific antibodies in the blood that are produced in response to infection. One of the first examples of FPA is the non-competitive test for detecting brucellosis in animals, which was followed by the development of other protocols for detecting various infections. Fluorescently labeled polysaccharides (in the case of brucellosis and salmonellosis) or specific peptides (in the case of tuberculosis and salmonellosis, etc.) can be used as biorecognition elements for detecting infections. The availability of new laboratory equipment and mobile devices for fluorescence polarization measurements outside the laboratory has stimulated the development of new fluorescence polarization assays (FPAs) and the emergence of commercial kits on the market for the detection of brucellosis, tuberculosis, and equine infectious anemia viruses. It has been shown that, in addition to antibodies, the FPA method can detect both viruses and nucleic acids. The development of more specific and sensitive biomarkers is essential for the diagnosis of infections and therapy monitoring. This review summarizes studies published between 2003 and 2023 that focus on the detection of infections using FPA. Furthermore, it demonstrates the potential for using new biorecognition elements (e.g., aptamers, proteins, peptides) and the combined use of FPA with new technologies, such as PCR and CRISPR/Cas12a systems, for detecting various infectious agents. Full article

The gut microbiome is shaped early in life by dietary and lifestyle factors. Specific compounds in the gut affect the growth of different bacterial species and the production of beneficial or harmful byproducts. Dysbiosis of the gut microbiome has been linked to various diseases resulting from the presence of harmful bacteria and their byproducts. Existing methods for detecting microbial species, such as microscopic observation and molecular biological techniques, are costly, labor-intensive, and require skilled personnel. Biosensors, which integrate a recognition element, transducer, amplifier, signal processor, and display unit, can convert biological events into electronic signals. This review provides a comprehensive and systematic survey of scientific publications from 2018 to June 2024, obtained from ScienceDirect, PubMed, and Scopus databases. The aim was to evaluate the current state-of-the-art and identify knowledge gaps in the application of aptamer biosensors for the determination of gut microbiota. A total of 13 eligible publications were categorized based on the type of study: those using microbial bioreceptors (category 1) and those using aptamer bioreceptors (category 2) for the determination of gut microbiota. Point-of-care biosensors are being developed to monitor changes in metabolites that may lead to disease. They are well-suited for use in the healthcare system and offer an excellent alternative to traditional methods. Aptamers are gaining attention due to their stability, specificity, scalability, reproducibility, low production cost, and low immunogenicity. While there is limited research on using aptamers to detect human gut microbiota, they show promise for providing accurate, robust, and cost-effective diagnostic methods for monitoring the gut microbiome. Full article

The early monitoring and inactivation of bacteria are of crucial importance in preventing the further spread of foodborne pathogens. Staphylococcus aureus (S. aureus), a prototypical foodborne pathogen, is widely present in the natural environment and has the capability to trigger a range of diseases at low concentrations. In this work, we designed Fe₃O₄@SiO₂-Au core–shell–satellite nanocomposites (NCs) modified with aptamer for efficient capture, high-sensitivity surface-enhanced Raman scattering (SERS) detection, and photothermal therapy (PTT) against S. aureus. Fe₃O₄@SiO₂-Au NCs with tunable Au nanocrystal nanogaps were prepared. By combining the finite-difference time-domain (FDTD) method and experimental results, we studied the electric field distribution of Fe₃O₄@SiO₂-Au under different Au nanogaps and ultimately obtained the optimal SERS substrate FSA-60. The modification of aptamer on the surfaces of FSA-60 could be used for the specific capture and selective detection of S. aureus, achieving a detection limit of as low as 50 cfu/mL. Furthermore, Apt-FSA-60 possessed excellent photothermal properties, demonstrating the strong photothermal killing ability against S. aureus. Therefore, Apt-FSA-60 is a promising high-sensitivity SERS substrate and efficient photothermal agent and is expected to be widely applied and promoted in future disease prevention and treatment. Full article

Massive use of antibiotics in veterinary medicine has led to their accumulation in meat and dairy products. Consumption of antibiotic-contaminated food can trigger the development of antibiotic-resistant bacteria, endangering human lives. Among antibiotics, the oxytetracycline (OTC) family of antibiotics is most widely used in veterinary medicine. Strict control of the antibiotics in food necessitates the development of fast and effective methods for OTC detection, for instance, in milk. One of the most promising approaches to OTC detection is based on the use of specially designed DNA aptamers. These DNA aptamers are relatively short, 15–60 bases, nucleotides folded in the solution in a 3D structure, forming a binding site for the target antibiotic. Aptamers can be chemically modified for attachment to sensor electrodes. In this work, we investigated the electrochemical detection of OTC using DNA aptamers specific to OTC that have been covalently immobilized onto the nanocomposite surface of a glassy carbon electrode with electrodeposited reduced graphene oxide and multiwalled carbon nanotubes. Differential pulse voltammetry, DPV, in the presentence of a ferri/ferrocyanide redox couple was used as an internal standard and to monitor the redox current. In the presence of OTC, the amplitude of DPV decreased, evidencing the blocking of charge transfer. After system optimization, we reached the limit of detection of 0.45 ng OTC/mL, which is 200 times lower than the maximum residue limit established by the European Commission of 100 ng OTC/mg. Full article

Bacterial lipopolysaccharides (LPSs) are important indicators of a bacteria presence in any samples. They can therefore be used for the detection of microbiological contamination in food and dairy products. We performed a comparative analysis of different bacterial models by the application of liposomes containing LPS from Salmonella enterica serotype typhimurium on the surface of an 11-mercaptoundecanoic acid (MUA) monolayer chemisorbed on the gold surface of quartz crystal. Using quartz crystal microbalance with dissipation monitoring (QCM-D), we were able to monitor the formation of the lectin, concanavalin A (ConA), layer on the MUA surface. We determined the optimal concentration of the ConA for the layer formation. ConA of 0.3 mg/mL was selected as the most suitable adsorption of liposomes containing LPS. Using the Sauerbrey equation, we calculated that approximately 1.13 × 10¹² ConA molecules per cm² was adsorbed on the MUA surface, which closely corresponds to the 1.19 × 10¹² molecules per cm² by theoretical models. Later, mixed LPS liposomes containing dipalmitoyl phosphatidyl choline (DPPC), dipalmitoyl phosphatidyl ethanolamine (DPPE) and cholesterol successfully interacted with the ConA layer, which resulted in a decrease in the resonant frequency and an increase in dissipation. We compared the adsorption of liposomes with different fractions of LPS and containing LPS from different bacteria. Lack of any LPS in liposomes caused weaker adsorption on the ConA layer. Liposomes containing 50% LPS caused the most prominent adsorption and were suitable for interaction with DNA aptamers specific to certain LPS. The addition of the aptamers to the surface of ConA covered by LPS-containing liposomes resulted in a decrease in resonant frequency and an increase in the dissipation. Using the Kelvin–Voigt viscoelastic model and multiharmonic response of acoustic sensors, we also determined changes in viscoelastic values of the molecular films during interaction with liposomes and the ConA layer. Full article

The rapid and sensitive detection of pathogenic bacteria is becoming increasingly important for the timely prevention of contamination and the treatment of infections. Biosensors based on nucleic acid aptamers, integrated with optical, electrochemical, and mass-sensitive analytical techniques, have garnered intense interest because of their versatility, cost-efficiency, and ability to exhibit high affinity and specificity in binding bacterial biomarkers, toxins, and whole cells. This review highlights the development of aptamers, their structural characterization, and the chemical modifications enabling optimized recognition properties and enhanced stability in complex biological matrices. Furthermore, recent examples of aptasensors for the detection of bacterial cells, biomarkers, and toxins are discussed. Finally, we explore the barriers to and discuss perspectives on the application of aptamer-based bacterial detection. Full article

Bacterial multidrug resistance poses a growing hazard to public health worldwide. Pseudomonas aeruginosa is a ubiquitous bacterial species that has been identified as the second most critical pathogen on the list of drug-resistant bacteria posing a huge threat to human health. P. aeruginosa is an opportunistic Gram-negative bacterium that rapidly produces multiple virulence factors that promote adhesion, host cell penetration, and pathogenicity. It possesses inherent resistance to multiple classes of antibiotics, evading antibiotic treatment by triggering the persister phenotype. Traditional approaches for detecting pathogens in laboratory and clinical settings primarily involve microbiological, nucleic acid-based, immunological, and sequencing techniques, among others. These procedures are time-consuming, necessitate advanced laboratory equipment and skilled staff, and incur large setup costs, rendering them unsuitable for on-the-spot detection of bacterial infections in settings with limited resources. Aptasensors utilize nucleic acid aptamers as bio-receptors to detect pathogens. They circumvent the existing limitations of conventional detection systems due to their sensitivity, versatility for modification, cost-efficiency, and ability to enable real-time detection. So far, as per the literature, three aptamers have been developed against P. aeruginosa using whole bacterial cells but with unreported binding sites. This study aims to identify highly specific aptamers that can bind specifically to P. aeruginosa using whole-bacterium SELEX at any stage of growth with an extensive study of binding site interaction and identification. The objective is to develop an aptamer-based kit enabling rapid point-of-care detection of P. aeruginosa in clinical and environmental samples. Full article

Early and accurate diagnoses of pathogenic microorganisms is essential to correctly identify diseases, treating infections, and tracking disease outbreaks associated with microbial infections, to develop precautionary measures that allow a fast and effective response in epidemics and pandemics, thus improving public health. Aptamers are a class of synthetic nucleic acid molecules with the potential to be used for medical purposes, since they can be directed towards any target molecule. Currently, the use of aptamers has increased because they are a useful tool in the detection of specific targets. We present a brief review of the use of aptamers to detect and identify bacteria or even some toxins with clinical importance. This work describes the advances in the technology of aptamers, with the purpose of providing knowledge to develop new aptamers for diagnoses and treatment of different diseases caused by infectious microorganisms. Full article

Optical fiber biosensors can be used to develop point-of-care tests (POCTs) for detecting viruses and bacteria in several matrices. In particular, the surface plasmon resonance (SPR) and localized SPR phenomena (LSPR) can be excited by exploiting low-cost and small-size optical fiber chips. Generally, SPR or LSPR sensors are realized using several kinds of modified optical fibers (silica, plastic, or specialty) or by exploiting other optical waveguides (e.g., slab, spoon-shaped waveguides, etc.). More specifically, optical fiber sensors can be classified as intrinsic or extrinsic. In the “optical fiber intrinsic sensors”, the sensing area is realized in the optical fiber directly, such as in the case of plasmonic platforms based on D-shaped plastic optical fibers (POFs), tapered optical fibers, U-bend POFs, or light-diffusing fibers (LDFs). By contrast, when an optical fiber is used as a mere waveguide allowing for the launch of light to the sensing region and its collection, it is defined as an extrinsic optical fiber sensor, like in the case of the plasmonic sensors realized by Cennamo et al. using POFs combined with spoon-shaped waveguides, 3D-printed platforms, bacterial cellulose waveguides, nanogratings, and InkJet-printed chips. To realize optical biosensor chips for the detection of viruses and bacteria, both intrinsic and extrinsic plasmonic POF sensors can be efficiently combined with receptors specific for membrane proteins, either biological (e.g., antibodies, aptamers, enzymes, etc.) or synthetic (e.g., molecularly imprinted polymers), to build groundbreaking POCTs. Full article

Aptamers have been spotlighted as promising bio-recognition elements because they can be tailored to specific target molecules, bind to targets with a high affinity and specificity, and are easy to chemically synthesize and introduce functional groups to. In particular, fluorescent aptasensors are widely used in biological applications to diagnose diseases as well as prevent diseases by detecting cancer cells, viruses, and various biomarkers including nucleic acids and proteins as well as biotoxins and bacteria from food because they have the advantages of a high sensitivity, selectivity, rapidity, a simple detection process, and a low price. We introduce screening methods for isolating aptamers with q high specificity and summarize the sequences and affinities of the aptamers in a table. This review focuses on aptamer-based fluorescence detection sensors for biological applications, from fluorescent probes to mechanisms of action and signal amplification strategies. Full article

Food and waterborne illnesses are still a major concern in health and food safety areas. Every year, almost 0.42 million and 2.2 million deaths related to food and waterborne illness are reported worldwide, respectively. In foodborne pathogens, bacteria such as Salmonella, Shiga-toxin producer Escherichia coli, Campylobacter, and Listeria monocytogenes are considered to be high-concern pathogens. High-concern waterborne pathogens are Vibrio cholerae, leptospirosis, Schistosoma mansoni, and Schistosima japonicum, among others. Despite the major efforts of food and water quality control to monitor the presence of these pathogens of concern in these kinds of sources, foodborne and waterborne illness occurrence is still high globally. For these reasons, the development of novel and faster pathogen-detection methods applicable to real-time surveillance strategies are required. Methods based on biosensor devices have emerged as novel tools for faster detection of food and water pathogens, in contrast to traditional methods that are usually time-consuming and are unsuitable for large-scale monitoring. Biosensor devices can be summarized as devices that use biochemical reactions with a biorecognition section (isolated enzymes, antibodies, tissues, genetic materials, or aptamers) to detect pathogens. In most cases, biosensors are based on the correlation of electrical, thermal, or optical signals in the presence of pathogen biomarkers. The application of nano and molecular technologies allows the identification of pathogens in a faster and high-sensibility manner, at extremely low-pathogen concentrations. In fact, the integration of gold, silver, iron, and magnetic nanoparticles (NP) in biosensors has demonstrated an improvement in their detection functionality. The present review summarizes the principal application of nanomaterials and biosensor-based devices for the detection of pathogens in food and water samples. Additionally, it highlights the improvement of biosensor devices through nanomaterials. Nanomaterials offer unique advantages for pathogen detection. The nanoscale and high specific surface area allows for more effective interaction with pathogenic agents, enhancing the sensitivity and selectivity of the biosensors. Finally, biosensors’ capability to functionalize with specific molecules such as antibodies or nucleic acids facilitates the specific detection of the target pathogens. Full article

Rikenella microfusus is an essential intestinal probiotic with great potential. The latest research shows that imbalance in the intestinal flora are related to the occurrence of various diseases, such as intestinal diseases, immune diseases, and metabolic diseases. Rikenella may be a target or biomarker for some diseases, providing a new possibility for preventing and treating these diseases by monitoring and optimizing the abundance of Rikenella in the intestine. However, the current monitoring methods have disadvantages, such as long detection times, complicated operations, and high costs, which seriously limit the possibility of clinical application of microbiome-based treatment options. Therefore, the intention of this study was to evolve an enriched aptamer library to be used for specific labeling of R. microfusus, allowing rapid and low-cost detection methods and, ultimately the construction of aptamer-based biosensors. In this study, we used Rikenella as the target bacterium for an in vitro whole Cell-SELEX (Systematic Evolution of Ligands by EXponential Enrichment) to evolve and enrich specific DNA oligonucleotide aptamers. Five other prominent anaerobic gut bacteria were included in this process for counterselection and served as control cells. The aptamer library R.m-R13 was evolved with high specificity and strong affinity (K_d = 9.597 nM after 13 rounds of selection). With this enriched aptamer library, R. microfusus could efficiently be discriminated from the control bacteria in complex mixtures using different analysis techniques, including fluorescence microscopy or fluorometric suspension assays, and even in human stool samples. These preliminary results open new avenues toward the development of aptamer-based microbiome bio-sensing applications for fast and reliable monitoring of R. microfusus. Full article