Detection Of Foodborne Bacteria Research Articles

Progress in Separation and Detection of Foodborne Bacteria for Food Safety

Progress in Separation and Detection of Foodborne Bacteria for Food Safety

A novel ensemble approach with deep transfer learning for accurate identification of foodborne bacteria from hyperspectral microscopy

The detection of foodborne bacteria is critical in ensuring both consumer safety and food safety. If these pathogens are not properly identified, it can lead to dangerous cross-contamination. One of the most common methods for classifying bacteria is through the examination of Hyperspectral microscope imaging (HMI). A widely used technique for measuring microbial growth is microscopic cell counting. HMI is a laborious and expensive process, producing voluminous data and needing specialized equipment, which might not be widely available. Machine learning (ML) methods are now frequently utilized to automatically interpret data from hyperspectral microscopy. The objective of our study is to devise a technique that employs deep transfer learning to address the challenge of limited data and utilizes four base classifiers - InceptionResNetV2, MobileNet, ResNet101V2, and Xception - to create an ensemble-based classification model for distinguishing live and dead bacterial cells of six pathogenic strains. In order to determine the optimal weights for the base classifiers, a Powell's optimization method was utilized in conjunction with a weighted average ensemble (WAVE) technique. We carried out an extensive experimental study to verify the efficiency of our proposed ensemble model on live and dead cell images of six different foodborne bacteria. In order to gain a better understanding of the regions, we performed a Grad-CAM analysis to explain the predictions made by our model. Through a series of experiments, our proposed framework has proven its capacity to effectively and precisely detect numerous bacterial pathogens. Specifically, it achieved a perfect identification rate of 100 % for Escherichia coli (EC), Listeria innocua (LI), and Salmonella Enteritidis (SE), while achieving rates of 96.30 % for Salmonella Typhimurium (ST), 87.13 % for Staphylococcus aureus (SA), and 94.12 % for Salmonella Heidelberg (SH). As a result, it can be considered as an effective tool for the identification of foodborne pathogens, due to its high level of efficiency.

Phage-Modified Clear Hydrogel for Simultaneous Detection of Multiple Bacteria.

The proliferation speed of live foodborne pathogens is fast. A small number of pathogens will have a great impact on food and the environment if positive samples are not detected timely. In this study, transparent porous hydrogel stir bars, modified by two different phages (corresponding to two different bacteria (Escherichia coli and Hafnia sp)), have been developed for rapid detection of foodborne bacteria. A large number of samples can be analyzed simultaneously with a small animal live imager device to screen out the positive samples, while an adenosine triphosphate (ATP) bioluminescence sensor can be used to quantify the number of bacteria in the positive samples. The phage has good specificity and capture ability to bacteria, which makes the method highly sensitive. In addition, the use of multiple phages also enables the method to detect multiple bacteria simultaneously. The three-dimensional structure of the hydrogel allows it to modify more phages, and its transparent nature also allows the inside bioluminescence to be detected. Both can enhance the sensitivity of the detection. Finally, the reagents needed for bioluminescence, such as d-luciferin, can also be preencapsulated in the hydrogel, thus simplifying the detection step. Under the best conditions, the detection range of the method is 102-108 CFU·mL-1, and the limit of detection is 30 CFU·mL-1 within 11 min. The test results of actual samples show that there is no difference between using the method developed through this study and the traditional plate counting method, but the detection time is greatly shortened.

Ultrasensitive electrochemical biosensor for bacteria detection based on Fe3O4@COF-AuNPs and trigging isothermal circular amplification

Foodborne bacterial infections pose serious threats to human health, leading to illnesses and even fatalities. The key to solve this problem is to develop fast and effective sensor for sensitive detection of bacteria. In this study, an electrochemical biosensor based on magnetic nanocomposite material Fe3O4@COF-AuNPs and trigging isothermal circular amplification (TICA) was constructed for detection of foodborne bacteria. In this design, a hairpin aptamer probe (HAP)-bound DNA monolayer was designed, serving as the substrate for the electrode to increase surface area. The Escherichia coli (E. coli) was selected as the target. The presence of E. coli caused the configuration change of the HAP and allowed the occurrence of TICA with the use of the Klenow Fragment polymerase (KF-Poly), thus the HAPs were away from the DNA monolayer. After adding the catalytic magnetic nanocomposite material Fe3O4@COF-AuNPs, it was immobilized on the DNA monolayer of the electrode through DNA complementary pairing and exhibited high electrocatalytic behavior towards the reduction of 4-nitrophenol. This enabled ultrasensitive detection of E. coli, with the detection limit (LOD) as low as 10 CFU/mL. The detection time was 1 h. This designed electrochemical sensor showed high selectivity, stability, and reproducibility, which offered a new avenue for detection of foodborne bacteria.

Functionalized carbon quantum dots fluorescent sensor array assisted by a machine learning algorithm for rapid foodborne pathogens identification

Food safety is a global concern, and conventional methods for detecting foodborne bacteria, such as polymerase chain reaction and enzyme-linked immunosorbent assay, often involve time-consuming processes, specialized equipment, or specific recognition of particular bacterial strains. There is an urgent need for more efficient and convenient detection methods for foodborne pathogens. This study addresses this need by introducing an easily constructed fluorescent sensor array for the identification of various foodborne bacteria. The sensor array comprises carbon quantum dots (CQDs) functionalized with ampicillin, polymyxin, and gentamicin, each exhibiting different affinities for binding with specific bacteria. Leveraging machine learning algorithms, the proposed sensor array enables rapid, accurate, and highly sensitive identification of foodborne pathogens. This approach offers a convenient solution for the development of rapid, accurate, and hypersensitive detection methods for multiple bioactive samples. In summary, the fluorescent sensor array in this study holds promise for advancing the field of foodborne bacteria detection.

Aptamer-mediated double strand displacement amplification with microchip electrophoresis for ultrasensitive detection of Salmonella typhimurium

Rapid and quantitative detection of foodborne bacteria is of great significance to public health. In this work, an aptamer-mediated double strand displacement amplification (SDA) strategy was first explored to couple with microchip electrophoresis (MCE) for rapid and ultrasensitive detection of Salmonella typhimurium (S. Typhimurium). In double-SDA, a bacteria-identified probe consisting of the aptamer (Apt) and trigger sequence (Tr) was ingeniously designed. The aptamer showed high affinity to the S. Typhimurium, releasing the Tr sequence from the probe. The released Tr hybridized with template C1 chain, initiating the first SDA to produce numerous output strands (OS). The second SDA process was induced with the hybridization of the liberated OS and template C2 sequence, generating a large number of reporter strands (RS), which were separated and quantified through MCE. Cascade signal amplification and rapid separation of nucleic acids could be realized by the proposed double-SDA method with MCE, achieving the limit of detection for S. typhimurium down to 6 CFU/mL under the optimal conditions. Based on the elaborate design of the probes, the double-SDA assisted MCE strategy achieved better amplification performance, showing high separation efficiency and simple operation, which has satisfactory expectation for bacterial disease diagnosis.

A portable CRISPR-Cas12a triggered photothermal biosensor for sensitive and visual detection of Staphylococcus aureus and Listeria monocytogenes

The detection of foodborne pathogens is crucial for ensuring the maintenance of food safety. In the present study, a portable CRISPR-Cas12a triggered photothermal biosensor integrating branch hybrid chain reaction (bHCR) and DNA metallization strategy for sensitive and visual detection of foodborne pathogens was proposed. The sheared probes were utilized to block the locker probes, which enabled preventing the assembly of bHCR in the absence of target bacteria, while target bacteria can activate the cleavage of sheared probes through CRISPR-Cas12a. Therefore, the locker probes functioned as initiating chains, triggering the formation of the branching double-stranded DNA consisting of H1, H2, and H3. The silver particles, which were in situ deposited on the DNA structure, functioned as a signal factor for conducting photothermal detection. Staphylococcus aureus and Listeria monocytogenes were selected as the foodborne pathogens to verify the analytical performance of this CRISPR-Cas12a triggered photothermal sensor platform. The sensor exhibited a sensitive detection with a low detection limit of 1 CFU/mL, while the concentration ranged from 100 to 108 CFU/mL. Furthermore, this method could efficiently detect target bacteria in multiple food samples. The findings demonstrate that this strategy can serve as a valuable reference for the development of a portable platform enabling quantitative analysis, visualization, and highly sensitive detection of foodborne bacteria.

Development of nucleic acid extraction and real-time recombinase polymerase amplification (RPA) assay integrated microfluidic biosensor for multiplex detection of foodborne bacteria

Development of nucleic acid extraction and real-time recombinase polymerase amplification (RPA) assay integrated microfluidic biosensor for multiplex detection of foodborne bacteria

Food-borne bacteria analysis using a diatomite bioinspired SERS platform.

Rapid and sensitive detection of food-borne bacteria has remained challenging over the past few decades. We propose a surface-enhanced Raman scattering sensing strategy based on a novel bioinspired surface-enhanced Raman scattering substrate, which can directly detect dye molecular residues and food-borne pathogen microorganisms in the environment. The surface-enhanced Raman scattering platform consists of a natural diatomite microporous array decorated with a metal-phenolic network that enables the in situ reduction of gold nanoparticles. The as-prepared nanocomposites display excellent surface-enhanced Raman scattering activity with the lowest limit of detection and the maximum Raman enhancement factor of dye molecules up to 10-11 M and 1.18 × 107, respectively. For food-borne bacterial detection, a diatomite microporous array decorated with a metal polyphenol network and gold nanoparticle-based surface-enhanced Raman scattering analysis is capable of distinguishing the biochemical fingerprint information of Staphylococcus aureus and Escherichia coli, indicating the great potential for strain identification.

Triazine-based covalent-organic framework embedded with cuprous oxide as the bioplatform for photoelectrochemical aptasensing Escherichia coli.

A superior photoelectrochemical (PEC) aptasensor was manufactured for the detection of Escherichia coli (E. coli) based on a hybrid of triazine-based covalent-organic framework (COF) and cuprous oxide (Cu2O). The COF synthesized using 1,3,5-tris(4-aminophenyl)-benzene (TAPB) and 1,3,5-triformylphloroglucinol (Tp) as building blocks acted as a scaffold for encapsulated Cu2O nanoparticles (denoted as Cu2O@TAPB-Tp-COF), which then was employed as the bioplatform for anchoring E. coli-targeted aptamer. Cu2O@Cu@TAPB-Tp-COF demonstrated enhanced separation of the photogenerated carriers and photoabsorption ability and boosted photoelectric conversion efficiency. The developed Cu2O@TAPB-Tp-COF-based PEC aptasensor exhibited a lower detection limit of 2.5CFUmL-1 toward E. coli within a wider range of 10CFUmL-1 to 1 × 104CFUmL-1 than most of reported aptasensors for determiningfoodborne bacteria, together with high selectivity, good stability, and superior ability and reproducibility. The recoveries of E. coli spiked into milk and bread samples ranged within 95.3-103.6% and 96.6-102.8%, accompanying with low RSDs of 1.37-4.48% and 1.74-3.66%, respectively. The present study shows a promising alternative for the sensitive detection of foodborne bacteria from complex foodstuffs and pathogenic bacteria-polluted environment.

Nanozyme-Mediated Catalytic Signal Amplification for Microfluidic Biosensing of Foodborne Bacteria.

Early detection of foodborne bacteria is urgently needed to ensure food quality and to avoid the outbreak of foodborne bacterial diseases. Here, a kind of metal-organic framework (Zr-MOF) modified with Pt nanoparticles (Pt-PCN-224) was designed as a peroxidase-like signal amplifier for microfluidic biosensing of foodborne bacteria. Taking Escherichia coli (E. coli) O157:H7 as a model, a linear range from 2.93 × 102 to 2.93 × 108 CFU/mL and a limit of detection of 2 CFU/mL were obtained. The whole detection procedure was integrated into a single microfluidic chip. Water, milk, and cabbage samples were successfully detected, showing consistency with the results of the standard culture method. Recoveries were in the range from 90 to 110% in spiked testing. The proposed microfluidic biosensor realized the specific and sensitive detection of E. coli O157:H7 within 1 h, implying broad prospects of MOF with biomimetic enzyme activities for biosensing.

Multivalent aptamer meshed open pore membrane and signal amplification for high-flux and ultra sensitive whole cell detection of E. coli O157:H7 in complex food matrices

We develop a multivalent aptamer meshed open pore membrane with a signal amplification method as a simple and robust platform for sensitive, robust, and high-flux detection of E. coli O157:H7 whole cells in food samples. Nylon open pore membranes are surface modified by multi-handled poly(amidoamine) dendrimers as templates that are subsequently functionalized with multiple branches of capturing RCA (cRCA). The resulting cRCA networks are composed of tandem repeating aptamers that can specifically recognize and capture E. coli O157:H7 cells. The captured cells are then conjugated with signaling RCA (sRCA) to amplify the fluorescent detection signal. Our results demonstrate that the cRCA-based aptamer meshwork significantly enhances the surface-capturing performance of E. coli O157:H7 cells while achieving high-fluxes. Moreover, the membrane surface modification using dendrimers renders the resulting membrane nonfouling thus significantly reducing the background noises by approximately 96 times while the sRCA signal intensification enhances the fluorescent detection signals by approximately 27 times. We show that the membranes exhibit excellent performances in detecting E.coli O157:H7 in milk and orange juice samples with limit of detection of 10 cells per 250 mL with excellent specificity, offering a robust and practical approach for rapid and sensitive detection of foodborne bacteria.

New Technique for Early Detection of Foodborne Bacteria

New Technique for Early Detection of Foodborne Bacteria

Fiber-Optic-Based Biosensor as an Innovative Technology for Point-of-Care Testing Detection of Foodborne Pathogenic Bacteria To Defend Food and Agricultural Product Safety.

Food safety is a concerning issue globally. Foodborne-pathogenic-bacteria-derived foodborne disease outbreaks have increased the threat to human health. The accurate and rapid detection of foodborne bacteria is of great significance for food safety. A fiber-optic-based biosensor has emerged as a powerful technique for the point-of-care testing of foodborne bacteria in food and agricultural products. This Perspective discusses the opportunities and challenges of fiber-optic-based biosensors for foodborne bacteria detection. The corresponding solution strategies to promote the application of this innovative technology in food and agricultural product detection for food safety and human health are also discussed and proposed.

Rapid and Online Detection of Foodborne Bacteria via a Novel Ultraviolet Photoionization Time-of-Flight Mass Spectrometry.

Foodborne bacteria are widespread contaminated sources of food; hence, the real-time monitoring of pathogenic bacteria in food production is important for the food industry. In this study, a novel rapid detection method based on microbial volatile organic compounds (MVOCs) emitted from foodborne bacteria was established by using ultraviolet photoionization time-of-flight mass spectrometry (UVP-TOF-MS). The results showed obvious differences of MVOCs among the five species of bacteria, and the characteristic MVOCs for each bacterium were selected by a feature selection algorithm. Online monitoring of MVOCs during bacterial growth displayed distinct metabolomic patterns of the five species. MVOCs were most abundant and varied among species during the logarithmic phase. Finally, MVOC production by bacteria in different food matrixes was explored. The machine learning models for bacteria cultured in different matrixes showed a good classification performance for the five species with an accuracy of over 0.95. This work based on MVOC analysis by online UVP-TOF-MS achieved effective rapid detection of bacteria and showed its great application potential in the food industry for bacterial monitoring.

Nanomaterial interfaces designed with different biorecognition elements for biosensing of key foodborne pathogens.

Foodborne diseases caused by pathogen bacteria are a serious problem toward the safety of human life in a worldwide. Conventional methods for pathogen bacteria detection have several handicaps, including trained personnel requirement, low sensitivity, laborious enrichment steps, low selectivity, and long-term experiments. There is a need for precise and rapid identification and detection of foodborne pathogens. Biosensors are a remarkable alternative for the detection of foodborne bacteria compared to conventional methods. In recent years, there are different strategies for the designing of specific and sensitive biosensors. Researchers activated to develop enhanced biosensors with different transducer and recognition elements. Thus, the aim of this study was to provide a topical and detailed review on aptamer, nanofiber, and metal organic framework-based biosensors for the detection of food pathogens. First, the conventional methods, type of biosensors, common transducer, and recognition element were systematically explained. Then, novel signal amplification materials and nanomaterials were introduced. Last, current shortcomings were emphasized, and future alternatives were discussed.

Salmonella typhimurium detection and ablation using OmpD specific aptamer with non-magnetic and magnetic graphene oxide

Foodborne diseases have increased in the last few years due to the increased consumption of packaged and contaminated food. Major foodborne bacteria cause diseases such as diarrhea, vomiting, and sometimes death. So, there is a need for early detection of foodborne bacteria as pre-existing detection techniques are time-taking and tedious. Aptamer has gained interest due to its high stability, specificity, and sensitivity. Here, aptamer has been developed against Salmonella Typhimurium through the Cell-Selex method, and to further find the reason for specificity and sensitivity, OmpD protein was isolated, and binding studies were done. Single molecular FRET experiment using aptamer and graphene oxide studies has also been done to understand the mechanism of FRET and subsequently used for target bacterial detection. Using this assay, Salmonella Typhimurium can be detected up to 10 CFU/mL. Further, Magnetic Graphene oxide was used to develop an assay to separate and ablate bacteria using 808 nm NIR where temperature increase was more than 60 °C within 30 s and has been shown by plating as well as a confocal live dead assay. Thus, using various techniques, bacteria can be detected and ablated using specific aptamer and Graphene oxide.

A Lab-on-a-Tube Biosensor Combining Recombinase-Aided Amplification and CRISPR-Cas12a with Rotated Magnetic Extraction for Salmonella Detection

Background: Foodborne pathogenic bacteria threaten worldwide public health, and simple bacterial detection methods are in urgent need. Here, we established a lab-on-a-tube biosensor for simple, rapid, sensitive, and specific detection of foodborne bacteria. Methods: A rotatable Halbach cylinder magnet and an iron wire netting with magnetic silica beads (MSBs) were used for simple and effective extraction and purification of DNA from the target bacteria, and recombinase-aided amplification (RAA) was combined with clustered regularly interspaced short palindromic repeats/CRISPR-associated proteins12a(CRISPR-Cas12a) to amplify DNA and generate fluorescent signal. First, 15 mL of the bacterial sample was centrifuged, and the bacterial pellet was lysed by protease to release target DNA. Then, DNA-MSB complexes were formed as the tube was intermittently rotated and distributed uniformly onto the iron wire netting inside the Halbach cylinder magnet. Finally, the purified DNA was amplified using RAA and quantitatively detected by the CRISPR-Cas12a assay. Results: This biosensor could quantitatively detect Salmonella in spiked milk samples in 75 min, with a lower detection limit of 6 CFU/mL. The fluorescent signal of 102 CFU/mL Salmonella Typhimurium was over 2000 RFU, while 104 CFU/mL Listeria monocytogenes, Bacillus cereus, and E. coli O157:H7 were selected as non-target bacteria and had signals less than 500 RFU (same as the negative control). Conclusions: This lab-on-a-tube biosensor integrates cell lysis, DNA extraction, and RAA amplification in one 15 mL tube to simplify the operation and avoid contamination, making it suitable for low-concentration Salmonella detection.

Coupling Bifunctional Nanozyme-Mediated Catalytic Signal Amplification and Label-Free SERS with Immunoassays for Ultrasensitive Detection of Pathogens in Milk Samples.

Rapid and sensitive detection of foodborne bacteria is of great significance in guaranteeing food safety and preventing foodborne diseases. A bifunctional Au@Pt core-shell nanozyme with excellent catalytic properties and high surface-enhanced Raman scattering (SERS) activity was developed for the highly sensitive detection of Salmonella typhimurium based on a label-free SERS strategy. The ultrathin Pt shell (about 1 nm) can catalyze Raman-inactive molecules into Raman-active reporters, greatly amplifying the amount of signal molecules. Moreover, the Au core serves as an active SERS substrate to enhance the signal of reporter molecules, further significantly improving the detection sensitivity. Benefiting from the excellent properties, such a bifunctional Au@Pt nanozyme was integrated with a magnetic immunoassay to construct a label-free SERS platform for the highly sensitive detection of S. typhi with a low detection limit of 10 CFU mL-1. Also, the Au@Pt-based SERS platform exhibited excellent selectivity and was successfully utilized for the detection of S. typhi in milk samples by a portable Raman spectrometer. Our demonstration of the bifunctional nanozyme-based SERS strategy provides an efficient pathway to improve the sensitivity of label-free SERS detection of pathogens and holds great promise in food safety, environmental analysis, and other biosensing fields.

Nanomaterial-based biosensors for the detection of foodborne bacteria: a review.

Ensuring food safety is a critical concern for the development and well-being of humanity, as foodborne illnesses caused by foodborne bacteria have increasingly become a major public health concern worldwide. Traditional food safety monitoring systems are expensive and time-consuming, relying heavily on specialized equipment and operations. Therefore, there is an urgent need to develop low-cost, user-friendly and highly sensitive biosensors for detecting foodborne bacteria. In recent years, the combination of nanomaterials with optical biosensors has provided a prospective future platform for the detection of foodborne bacteria. By harnessing the unique properties of nanomaterials, such as their high surface area-to-volume ratio and exceptional sensitivity, in tandem with the precision of optical biosensing techniques, a new prospect has opened up for the rapid and accurate identification of potential bacterial contaminants in food. This review focuses on recent advances and new trends of nanomaterial-based biosensors for the detection of foodborne pathogens, which mainly include noble metal nanoparticles (NMPs), metal organic frameworks (MOFs), graphene nanomaterials, quantum dot (QD) nanomaterials, upconversion fluorescent nanomaterials (UCNPs) and carbon dots (CDs). Additionally, we summarized the research progress of color indicators, nanozymes, natural enzyme vectors and fluorescent dye biosensors, focusing on the advantages and disadvantages of nanomaterial-based biosensors and their development prospects. This review provides an outlook on future technological directions and potential applications to help identify the most promising areas of development in this field.