Foodborne pathogens cause many diseases and significantly impact human health and the economy. Foodborne pathogens mainly include Salmonella spp., Escherichia coli, Staphylococcus aureus, Shigella spp., Campylobacter spp. and Listeria monocytogenes, which are present in agricultural products, dairy products, animal-derived foods and the environment. Various pathogens in many different types of food and water can cause potentially life-threatening diseases and develop resistance to various types of antibiotics. The harm of foodborne pathogens is increasing, necessitating effective and efficient methods for early monitoring and detection. Traditional methods, such as real-time polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA) and culture plate, are time-consuming, labour-intensive and expensive and cannot satisfy the demands of rapid food testing. Therefore, new fast detection methods are urgently needed. Electrochemical biosensors provide consumer-friendly methods to quickly detect foodborne pathogens in food and the environment and achieve extensive accuracy and reproducible results. In this paper, by focusing on various mechanisms of electrochemical transducers, we present a comprehensive overview of electrochemical biosensors for the detection of foodborne pathogens. Furthermore, the review introduces the hazards of foodborne pathogens, risk analysis methods and measures of control. Finally, the review also emphasizes the recent research progress and solutions regarding the use of electrochemical biosensors to detect foodborne pathogens in food and the environment, evaluates limitations and challenges experienced during the development of biosensors to detect foodborne pathogens and discusses future possibilities. 

Recent Advances in Electrochemical Biosensors for the Detection of Foodborne Pathogens: Current Perspective and Challenges

This article presents a review of recent advancements in the utilization of NAA-based techniques for detecting foodborne pathogens in food products, focusing on studies conducted within the past three years. This review revealed that recent research efforts have primarily aimed at enhancing sensitivity and specificity by improving sample pre-treatment/preparation, DNA isolation, and readout methods. Isothermal-based amplification methods, such as LAMP, RPA, RAA, and RCA, have emerged as promising approaches, providing rapid results within one h and often demonstrating comparable or superior sensitivity to conventional or qPCR methods. However, the attention paid to specific pathogens varies, with Salmonella, Listeria, E. coli, and V. parahaemolyticus receiving more focus than norovirus and other similar pathogens. NAA-based methods have the potential to significantly contribute to food safety and public health protection. However, further advancements are necessary to fully realize their benefits. 

Rapid detection methods for foodborne pathogens based on nucleic acid amplification: Recent advances, remaining challenges, and possible opportunities

An accurate and ultrasensitive ratiometric electrochemical aptasensor for determination of ochratoxin A based on catalytic hairpin assembly

Foodborne Disease Symptoms, Diagnostics, and Predictions Using Artificial Intelligence-Based Learning Approaches: A Systematic Review

The global food demand is expected to increase in the coming years, along with challenges around climate change and food security. Concomitantly, food safety risks, particularly those related to bacterial pathogens, may also increase. Thus, the food sector needs to innovate to rise to this challenge. Here, we discuss recent advancements in molecular techniques that can be deployed within various foodborne bacteria surveillance systems across food settings. To start with, we provide updates on nucleic acid-based detection, with a focus on polymerase chain reaction (PCR)-based technologies and loop-mediated isothermal amplification (LAMP). These include descriptions of novel genetic markers for several foodborne bacteria and progresses in multiplex PCR and droplet digital PCR. The next section provides an overview of the development of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) proteins systems, such as CRISPR-Cas9, CRISPR-Cas12a, and CRISPR-Cas13a, as tools for enhanced sensitive and specific detection of foodborne pathogens. The final section describes utilizations of whole genome sequencing for accurate characterization of foodborne bacteria, ranging from epidemiological surveillance to model-based predictions of bacterial phenotypic traits through genome-wide association studies or machine learning.

https://onlinelibrary.wiley.com/doi/pdfdirect/10.1111/1541-4337.13153

Molecular detection and characterization of foodborne bacteria: Recent progresses and remaining challenges.

An ultrasensitive and specific ratiometric electrochemical biosensor based on SRCA-CRISPR/Cas12a system for detection of Salmonella in food

Aflatoxin B1 is a potent carcinogen widely found in agricultural products and feed. Therefore, investigating the development of convenient, sensitive, and precise techniques for detecting AFB1 is critical. This work proposes a ratiometric electrochemical aptasensor based on DNAzyme-driven tripedal DNA walker for enzyme-free AFB1 detection. The design of multiple legs reduces the moving time on the electrode surface and enhances the value of signal response. In this approach, the tagging of ferrocene and methylene blue as signal switch molecules diminishes the impact of interferences and significantly enhances the reproducibility (RSD: 2.825%). Meanwhile, the tripedal DNA walker and DNAzyme dual signal amplification enabled the sensor to detect AFB1 traces in cereals.We achieved trace detection of AFB1 with a detection limit of 61 fg/mL and a wide linear range from 0.0001 to 10 ng/mL. Furthermore, the aptasensor success rate was comparable to that of HPLC-PMP. The results demonstrate enormous potential for detection. Furthermore, the method can detect various targets by altering the target aptamer, displaying attractive scalability. This suggests its tremendous prospective in detecting mycotoxins in food in the future. 

DNAzyme-driven tripedal DNA walker for ratiometric electrochemical aptasensor ultrasensitive detection of aflatoxin B1

Saltatory rolling circle amplification assay coupled with photosensitization method for rapid and sensitive detection of Salmonella in food

In response to the potential controversies on increasing circulation of genetically modified organisms (GMOs) in the global markets, the rapid and sensitive detection methods are essential for monitoring of GMOs to maintain consumers' rights. Herein, we first developed a rapid technique for the real-time and visual on-site detection of the genetically modified crops (specifically targeting CP4-EPSPS gene) by using single primer isothermal amplification (SPIA) combined with SYBR Green II. 19 GM crop events and 12 non-GM crop events were used to assess the specificity of the developed SPIA assay, which presented excellent specificity. The limit of detection of the developed SPIA method was as low as 4 copies/μL, which was better than that of the real-time PCR method (20 copies/μL). The SPIA method was further used to assess 30 of actual samples, and the experimental results revealed that 3 positive samples were found, which were in good agreement with those of the real-time PCR methods. Overall, the major advantages of the developed SPIA over real-time PCR are the lower LOD, simplicity, and inexpensiveness. The developed SPIA method provides a potential alternative for the real-time and visual on-site detection of GMOs, which also accelerates the development of highly sensitive nucleic acid testing techniques. 

Single primer isothermal amplification for real-time and visual on-site detection of genetically modified crops by recognizing CP4-EPSPS gene

Rapid and visual detection of viable Staphylococcus aureus in pork and pork products by PMA and saltatory rolling circle amplification

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