Detection Of Foodborne Pathogenic Bacteria Research Articles

A novel MNPs@GO biosensor- integrated upconversion fluorescence cuvette for simultaneous detection of Salmonella typhimurium and Staphylococcus aureus

An upconversion fluorescence biosensor was developed and employed in a self-designed cuvette engineered for the detection of Salmonella typhimurium and Staphylococcus aureus in chicken samples. The cuvette consisted of a hollow cylinder filled with alendronic acid-modified upconversion nanoparticles (ADA@UCNPs) and solid quartz cube blocks and could provide a stable and reusable fluorescent donor. In addition, the fluorescence inner filter mechanism (IEF) between two fluorescence quenchers (3,3′,5,5′-tetramethylbenzidine oxide (oxTMB) and 5-bromo-4-chloro-3-indolyl phosphate/nitroblue tetrazole (oxBCIP/NBT)) and ADA@UCNPs in the cuvette could be triggered. After sample injection, bacteria-targeting aptamers labeled with horseradish peroxidase (HRP) or alkaline phosphatase (AP) fell off the magnetic graphene oxide (MNPs@GO), resulting in the blockage of the enzymatic reaction. This led to an increase in the yellow-green upconversion fluorescence peaks at 547 nm and 658 nm. The whole detection process could be accomplished within 40 min and required the minimum amounts of ADA@UCNPs and MNPs@GO. The limit of detection was as low as 14 CFU/mL for Salmonella typhimurium and 33 CFU/mL for Staphylococcus aureus. To sum up, the upconversion fluorescence biosensor holds a potential in rapid analysis and detection of foodborne pathogenic bacteria.

Investigating enzyme kinetics and fluorescence sensing strategy of CRISPR/Cas12a for foodborne pathogenic bacteria

Foodborne pathogenic bacteria are widespread in various foods, whose cross-contamination and re-contamination are critical influences on food safety. Rapid, accurate, and sensitive detection of foodborne pathogenic bacteria remains a topic of concern. CRISPR/Cas12a can recognize double-stranded DNA directly, showing great potential in nucleic acid detection. However, few studies have investigated the cleavage properties of CRISPR/Cas12a. In this study, the trans-cleavage properties of LbCas12a and AsCas12a were investigated to construct the detection methods for foodborne pathogenic bacteria. The highly sensitive fluorescent strategies for foodborne pathogens were constructed by analyzing the cleavage rates and properties of substrates at different substrate concentrations. Cas12a was activated in the presence of foodborne pathogenic target sequence was present, resulting in the cleavage of a single-stranded reporter ssDNA co-labelled by fluorescein quencher and fluorescein. The sensitivity and specificity of the Cas12a fluorescent strategy was investigated with Salmonella and Staphylococcus aureus as examples. The results showed that AsCas12a was slightly more capable of trans-cleavage than LbCas12a. The detection limits of AsCas12a for Salmonella and Staphylococcus aureus were 24.9 CFU mL−1 and 1.50 CFU mL−1, respectively. In all the seven bacteria, Staphylococcus aureus and Salmonella were accurately discriminated. The study provided a basis for constructing and improving the CRISPR/Cas12a fluorescence strategies. The AsCas12a-based detection strategy is expected to be a promising method for field detection.

Double phage displayed peptides co-targeting-based biosensor with signal enhancement activity for colorimetric detection of Staphylococcus aureus

The development of simple, fast, sensitive, and specific strategies for the detection of foodborne pathogenic bacteria is crucial for ensuring food safety and promoting human health. Currently, detection methods for Staphylococcus aureus still suffer from issues such as low specificity and low sensitivity. To address this problem, we proposed a sensitivity enhancement strategy based on double phage-displayed peptides (PDPs) co-targeting. Firstly, we screened two PDPs and analyzed their binding mechanisms through fluorescent localization, pull-down assay, and molecular docking. The two PDPs target S. aureus by binding to specific proteins on its outer membrane. Based on this phenomenon, a convenient and sensitive double PDPs colorimetric biosensor was developed. Double thiol-modified phage-displayed peptides (PDP-SH) enhance the aggregation of gold nanoparticles (AuNPs), whereas the specific interaction between the double PDPs and bacteria inhibits the aggregation of AuNPs, resulting in an increased visible color change before and after the addition of bacteria. This one-step colorimetric approach displayed a high sensitivity of 2.35 CFU/mL and a wide detection range from 10–2 × 108 CFU/mL. The combination with smartphone-based image analysis improved the portability of this method. This strategy achieves the straightforward, highly sensitive and portable detection of pathogenic bacteria.

Rapid, on-site and yes-or-no detection of Salmonella typhimurium in foods using Argonaute-enabled assay

Early detection and control of foodborne diseases are crucial for ensuring food safety and consumer health. Therefore, there is a need for faster and more sensitive detection methods. We developed a rapid, portable and visible lateral flow biosensor (LFB) for foodborne pathogenic bacteria detection powered by a Pyrococcus furiosus Argonaute nuclease (PfAgo), named as PfAgo-LFB. LAMP (Loop-mediated isothermal amplification) was used to amplify the target nucleic acid. By using PfAgo as the “lighting up” technology, we can overcome the limitations of false-positive results and lack of effective endpoint monitoring in LAMP, greatly enhancing its applicability. It turned out that PfAgo-LFB was able to provide yes-or-no (positive or negative) results with a limit of detection (LOD) as low as 1 CFU/mL for S. typhimurium and the detection range was between 100 to 108 CFU/mL. It was able to secure a sample-to-answer test for contaminated food samples in less than 45 min and demonstrated satisfactory selectivity. In summary, we designed a PfAgo-LFB that rendered a novel, rapid, accurate, easy-to-deploy, on-site and yes or no detection of S. typhimurium, which could be generalized for building a “one-for-all” biosensing platform.

E. coli O157:H7 Detection Using Surface Plasmon Resonance Based Biosensor

The detection of foodborne pathogenic bacteria remains a significant challenge, and the need for fast and sensitive detection methods is becoming increasingly important. Escherichia coli is a prevalent bacteria associated with foodborne illness, and this study aimed to evaluate the ability of a surface plasmon resonance (SPR) based biosensor to detect E. coli O157:H7 at low levels in pure culture and artificially contaminated bay leaves (Laurus nobilis) using different injection methods. To develop a biological sensing surface, the sensor surface was functionalized with 3-aminopropyltriethoxysilane (APTES), and polyclonal antibodies were immobilized on the surface for bacteria detection. Bacterial attachment to the antibodies resulted in a change in resonance angle. The biosensor was able to discriminate between cellular concentrations of 103 to 107 CFU/mL and showed potential in detecting different pathogens in various food samples. Before the SPR detection, the sample preparation step was optimized to ensure complex food matrices were suitable for SPR analysis. The results suggest that the SPR based biosensor is a promising tool for the rapid detection of foodborne pathogens in complex food matrices.

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.

Lateral Flow Strip Biosensors for Foodborne Pathogenic Bacteria via Direct and Indirect Sensing Strategies: A Review.

Rapid and sensitive detection of foodborne pathogenic bacteria is particularly important for the prevention and control of foodborne diseases. The lateral flow strip biosensor (LFSB) is one of the most promising point-of-care detection tools and has been widely used in food safety monitoring. This review introduces recent advances in the detection of foodborne pathogenic bacteria using LFSBs. According to different bacterial biomarkers, we summarize the direct and indirect sensing strategies of bacterial LFSBs. The direct sensing strategies for whole bacterial cells are divided into antibodies, antibody alternatives, and label-free according to the recognition elements. The indirect sensing strategies refer to the detection of bacterial nucleic acids and metabolites. Next, we compare and discuss the applications of direct and indirect sensing strategies. Finally, the existing challenges, future perspectives, and development directions are discussed, which will facilitate the theoretical innovation and practical application for bacterial LFSBs.

Ag+-gated peroxidase activity of gold nanoparticles for sensitive detection of Escherichia coli

Escherichia coli (E. coli) is one of the most ubiquitous foodborne pathogens that can cause infections and threaten human health. Herein, a colorimetric method for sensitive detection of E. coli was established by using enzyme-nanozyme cascade reaction for signal amplification. Gold nanoparticles (AuNPs) are well-known nanozymes due to their high peroxidase-like activity. When the dense cetyltrimethylammonium bromide (CTAB) membrane on the surfaces of AuNPs kept the substrate away from AuNPs, the peroxidase activity of AuNPs was inhibited. However, the CTAB membrane could be disrupted by Ag+, resulting in enhanced peroxidase activity of AuNPs. When E. coli was present, the enzyme-nanozyme cascade reaction was initiated. The substrate p-aminophenyl β-D-galactopyranoside (PAPG) was hydrolyzed to the reductive p-aminophenol (PAP) by beta-galactosidase (β-gal) in E. coli, reducing Ag+ to Ag. Consequently, CTAB-AuNPs remained weak peroxidase activity and could not catalyze the H2O2-mediated oxidation of TMB. As the amount of E. coli increased, the absorbance of TMB decreased along with a color change from deep blue to pink. The absorbance intensity displayed a linear dependence on E. coli from 1.0 × 102 to 1.0 × 109 CFU mL−1. Therefore, the proposed method holds good prospects in foodborne pathogenic bacteria detection.

Argonaute-triggered visual and rebuilding-free foodborne pathogenic bacteria detection

Foodborne pathogenic bacteria are recognized as the main causes of microbial contamination in food safety. Early screening and ultrasensitive detection of foodborne pathogenic bacteria is critical procedure to guarantee food safety. Argonaute is emerging as a new tool for detection owing to the programmability and high specificity. We reported a Novel and One-step cleavage method based on Argonaute by integrating Tag-specific primer extension and Exonuclease I (Exo I) for the first time, termed as NOTE-Ago. In this method, the invA of Salmonella typhi and nuc gene of Staphylococcus aureus were amplified using Tag-specific primer and the remaining primers were digested by Exo I. Then amplicons were served as the guide DNA for PfAgo. Consequently, the fluorophore-quencher reporter could be cleaved via PfAgo, resulting in changes in fluorescent intensity. With this strategy, target nucleic acid could be dexterously converted into fluorescent signals. The NOTE-Ago assay could detect 1 CFU/mL with a dynamic range from 1 to 108 CFU/mL. The satisfactory selectivity of NOTE-Ago assay further facilitated its application for detecting S. typhi- and S. aureus-contaminated food samples. This work enriches the toolbox of Argonaute-based detection and provides a one-step cleavage and rebuilding-free method for ultrasensitive detection of bacteria.

Capture, sterilization and determination platform of foodborne pathogen based on Halbach ring-mediated magnetic filter

Herein, an ingenious Halbach ring-mediated magnetic filter (HRMF) for capture, sterilization, and detection of foodborne pathogenic bacteria is developed. A composite material with magnetic nanoparticles (MNP) as the core and coated with polydopamine (PDA) and polyethylenimine (PEI) is synthesized, namely MNP@PDA@PEI. Under a Halbach ring, MNP@PDA@PEI can form chains to generate HRMF. Due to the positively charged of PEI, the HRMF could capture the negatively charged bacteria from solution via electrostatic adsorption. Within the HRMF, sterilization performance could be achieved due to the photothermal effect of MNP@PDA@PEI after 5 min of near-infrared light irradiation. More importantly, the number of bacteria could be calculated by bioluminescence method for measuring ATP, which are released from bacteria after photothermal sterilization. Listeria monocytogenes is chosen as a typical bacterium to evaluate the function of HRMF. The time of whole process has been greatly reduced to 38 min. The capture efficiency is 98.63%, and sterilization rate can achieve 99.96% in real samples. The limit of detection is 10.04 CFU/mL, representing a 10-fold decrease compared with bioluminescence method alone. The proposed platform might open a new way for on-site real-time detection of foodborne pathogens, and hold potential to nondestructive sterilization in the food processing industry.

One-Pot Synthesis of Enzyme and Antibody/CaHPO4 Nanoflowers for Magnetic Chemiluminescence Immunoassay of Salmonella enteritidis.

In this study, through a bioinspired strategy, the horseradish peroxidase (HRP) and antibody (Ab) were co-embedded into CaHPO4 to prepare HRP-Ab-CaHPO4 (HAC) bifunctional hybrid nanoflowers by one-pot mild coprecipitation. The as-prepared HAC hybrid nanoflowers then were utilized as the signal tag in a magnetic chemiluminescence immunoassay for application in the detection of Salmonella enteritidis (S. enteritidis). The proposed method exhibited excellent detection performance in the linear range of 10-105 CFU/mL, with the limit of detection (LOD) of 10 CFU/mL. This study indicates great potential in the sensitive detection of foodborne pathogenic bacteria in milk with this new magnetic chemiluminescence biosensing platform.

Molecular Targets for Foodborne Pathogenic Bacteria Detection.

The detection of foodborne pathogenic bacteria currently relies on their ability to grow on chemically defined liquid and solid media, which is the essence of the classical microbiological approach. Such procedures are time-consuming and the quality of the result is affected by the selectivity of the media employed. Several alternative strategies based on the detection of molecular markers have been proposed. These markers may be cell constituents, may reside on the cell envelope or may be specific metabolites. Each marker provides specific advantages and, at the same time, suffers from specific limitations. The food matrix and chemical composition, as well as the accompanying microbiota, may also severely compromise detection. The aim of the present review article is to present and critically discuss all available information regarding the molecular targets that have been employed as markers for the detection of foodborne pathogens. Their strengths and limitations, as well as the proposed alleviation strategies, are presented, with particular emphasis on their applicability in real food systems and the challenges that are yet to be effectively addressed.

The Detection of Foodborne Pathogenic Bacteria in Seafood Using a Multiplex Polymerase Chain Reaction System

Multiplex polymerase chain reaction (PCR) assays are mainly used to simultaneously detect or identify multiple pathogenic microorganisms. To achieve high specificity for detecting foodborne pathogenic bacteria, specific primers need to be designed for the target strains. In this study, we designed and achieved a multiplex PCR system for detecting eight foodborne pathogenic bacteria using specific genes: toxS for Vibrio parahaemolyticus, virR for Listeria monocytogenes, recN for Cronobacter sakazakii, ipaH for Shigella flexneri, CarA for Pseudomonas putida, rfbE for Escherichia coli, vvhA for Vibrio vulnificus, and gyrB for Vibrio alginolyticus. The sensitivity of the single system in this study was found to be 20, 1.5, 15, 15, 13, 14, 17, and 1.8 pg for V. parahaemolyticus, L. monocytogenes, E. coli O157:H7, C. sakazakii, S. flexneri, P. putida, V. vulnificus, and V. alginolyticus, respectively. The minimum detection limit of the multiplex system reaches pg/μL detection level; in addition, the multiplex system exhibited good specificity and stability. Finally, the assays maintained good specificity and sensitivity of 104 CFU/mL for most of the samples and we used 176 samples of eight aquatic foods, which were artificially contaminated to simulate the detection of real samples. In conclusion, the multiplex PCR method is stable, specific, sensitive, and time-efficient. Moreover, the method is well suited for contamination detection in these eight aquatic foods and can rapidly detect pathogenic microorganisms.

Recent Progress in Spectroscopic Methods for the Detection of Foodborne Pathogenic Bacteria.

Detection of foodborne pathogens at an early stage is very important to control food quality and improve medical response. Rapid detection of foodborne pathogens with high sensitivity and specificity is becoming an urgent requirement in health safety, medical diagnostics, environmental safety, and controlling food quality. Despite the existing bacterial detection methods being reliable and widely used, these methods are time-consuming, expensive, and cumbersome. Therefore, researchers are trying to find new methods by integrating spectroscopy techniques with artificial intelligence and advanced materials. Within this progress report, advances in the detection of foodborne pathogens using spectroscopy techniques are discussed. This paper presents an overview of the progress and application of spectroscopy techniques for the detection of foodborne pathogens, particularly new trends in the past few years, including surface-enhanced Raman spectroscopy, surface plasmon resonance, fluorescence spectroscopy, multiangle laser light scattering, and imaging analysis. In addition, the applications of artificial intelligence, microfluidics, smartphone-based techniques, and advanced materials related to spectroscopy for the detection of bacterial pathogens are discussed. Finally, we conclude and discuss possible research prospects in aspects of spectroscopy techniques for the identification and classification of pathogens.

Simultaneous detection of foodborne pathogenic bacteria in milk by fluorescence immunoassay

Rapid, sensitive and simultaneous detection of multiple bacteria in foodborne is still a major challenge in public health field. Here, a fluorescence immunoassay that can achieve high-throughput detection of three Gram-positive foodborne pathogenic bacteria simultaneously was proposed. Vancomycin and bovine serum albumin conjugate (Van-BSA) was immobilized on a polycarbonate chip to capture three Gram-positive foodborne pathogenic bacteria, Staphylococcus aureus (S. aureus), Bacillus cereus (B. cereus) and Listeria monocytogenes (L. monocytogenes). CdSe/ZnS quantum dot modified antibodies (Ab-QD) were prepared by carbodiimide coupling chemistry. Due to the affinity reaction between antibodies and proteins on the bacterial surface, the simultaneous detection of multiple Gram-positive bacteria was achieved by monitoring the fluorescence signal of quantum dot by a portable microfluidic chip analyzer. Under optimal conditions, low detection limits was 18 CFU/well, 3 CFU/well and 36 CFU/well for S. aureus, B. cereus and L. monocytogenes, respectively. With satisfactory accuracy and precision, the proposed fluorescence immunoassay holds good prospects to detect pathogens in real food samples.

Research progress on detection techniques for point-of-care testing of foodborne pathogens.

The global burden of foodborne disease is enormous and foodborne pathogens are the leading cause of human illnesses. The detection of foodborne pathogenic bacteria has become a research hotspot in recent years. Rapid detection methods based on immunoassay, molecular biology, microfluidic chip, metabolism, biosensor, and mass spectrometry have developed rapidly and become the main methods for the detection of foodborne pathogens. This study reviewed a variety of rapid detection methods in recent years. The research advances are introduced based on the above technical methods for the rapid detection of foodborne pathogenic bacteria. The study also discusses the limitations of existing methods and their advantages and future development direction, to form an overall understanding of the detection methods, and for point-of-care testing (POCT) applications to accurately and rapidly diagnose and control diseases.

Recognition of Foodborne Pathogens Terahertz Spectrum Based on Convolutional Neural Network

Foodborne pathogenic bacteria detection is an important work, the traditional laboratory identification method, will cause irreversible damage to the sample, and the detection cycle is long. Compared with traditional biological detection methods, the greatest advantage of terahertz spectroscopy detection is that it can do non-destructive detection of samples, and terahertz spectroscopy technology, combined with computer systems, can reduce analysis time, improve recognition accuracy, and maximize the extraction and mining of effective information in the spectrum. This paper establishes an intelligent foodborne pathogenic bacteria terahertz spectroscopic recognition model, The results showed that the recognition accuracy of pathogens was 99.76%.

State of the art: Lateral flow assays toward the point-of-care foodborne pathogenic bacteria detection in food samples.

Diverse chemicals and some physical phenomena recently introduced in nanotechnology have enabled scientists to develop useful devices in the field of food sciences. Concerning such developments, detecting foodborne pathogenic bacteria is now an important issue. These kinds of bacteria species have demonstrated severe health effects after consuming foods and high mortality related to acute cases. The most leading path of intoxication and infection has been through food matrices. Hence, quick recognition of foodborne bacteria agents at low concentrations has been required in current diagnostics. Lateral flow assays (LFAs) are one of the urgent and prevalently applied quick recognition methods that have been settled for recognizing diverse types of analytes. Thus, the present review has stressed on latest developments in LFAs-based platforms to detect various foodborne pathogenic bacteria such as Salmonella, Listeria, Escherichia coli, Brucella, Shigella, Staphylococcus aureus, Clostridium botulinum, and Vibrio cholera. Proper prominence has been given on exactly how the labels, detection elements, or procedures have affected recent developments in the evaluation of diverse bacteria using LFAs. Additionally, the modifications in assays specificity and sensitivity consistent with applied food processing techniques have been discussed. Finally, a conclusion has been drawn for highlighting the main challenges confronted through this method and offered a view and insight of thoughts for its further development in the future.

Recent advancements in microfluidic chip biosensor detection of foodborne pathogenic bacteria: a review.

Foodborne diseases caused by pathogenic bacteria pose a serious threat to human health. Early and rapid detection of foodborne pathogens is an urgent task for preventing disease outbreaks. Microfluidic devices are simple, automatic, and portable miniaturized systems. Compared with traditional techniques, microfluidic devices have attracted much attention because of their high efficiency and convenience in the concentration and detection of foodborne pathogens. This article firstly reviews the bio-recognition elements integrated on microfluidic chips in recent years and the progress of microfluidic chip development for pathogen pretreatment. Furthermore, the research progress of microfluidic technology based on optical and electrochemical sensors for the detection of foodborne pathogenic bacteria is summarized and discussed. Finally, the future prospects for the application and challenges of microfluidic chips based on biosensors are presented.Graphical abstract

CRISPR/Cas12a and immuno-RCA based electrochemical biosensor for detecting pathogenic bacteria

Scheme 1 Schematic illustration of immuno-RCA and Cas12a based electrochemical biosensor for quantitative analysis of E. coli O157:H7. • An electrochemical biosensor based on CRISPR was established to detect bacteria. • The electrochemical biosensor can be used in the detection of actual samples. • The immuno-RCA was used to amplify the activity of CRISPR/Cas12a system. • Have a wide detection range from 10 to 10 7 CFU•mL −1 , with a LOD of 10 CFU·mL −1 . Rapid, simple, and sensitive detection of food-borne pathogenic bacteria is extremely important for preventing and controlling food-borne diseases. Herein, an electrochemical biosensor based on CRISPR/Cas12a combined with immuno-rolling circle amplification (immuno-RCA) was developed for detecting the pathogenic bacterium, Escherichia coli O157:H7. Based on a sandwich-type immunoassay on magnetic beads, immuno-RCA generated long single-stranded DNA with abundant E. coli O157:H7-specific aptamers and target repeated sequences, which allowed recognition and binding of E. coli O157:H7 and complex formation by CRISPR/Cas12a and crRNA, respectively. Stimulation of Cas12a produced trans- cleavage activity toward a non-specific methylene blue labeled DNA hairpin probe on the electrode surface. In the presence of E. coli O157:H7, the CRISPR/Cas12a non-specific trans- cleavage activity was triggered, the hairpin DNA on the Au electrode was cleaved, and the peak current was altered. Under optimal conditions, the developed biosensor presented a broad dynamic detection range from 10 to 10 7 CFU·mL −1 , with a detection limit of 10 CFU·mL −1 . Moreover, the biosensor did not exhibit any cross-reactivity with other non-target bacteria. These results revealed that the developed biosensor is a simple, sensitivity, and specific platform for E. coli O157:H7 detection.