Rapid detection of intact pathogens on a microfluidic device by combining immunomagnetic separation and viability dyes

Recent advances in microfluidic devices for foodborne pathogens detection

Microfluidic devices for sample preparation and rapid detection of foodborne pathogens

Rapid detection of pathogens and assessment of antimicrobial susceptibility is of great importance for public health, especially in resource-limiting regions. Herein, we developed a rapid, portable, and universal detection method for bacteria using AgNPs-invertase complexes and the personal glucose meter (PGM). In the presence of bacteria, the invertase could be released from AgNPs-invertase complexes where its enzyme activity of invertase was inhibited. Then, the enzyme activity of invertase was restored and could convert sucrose into glucose measured by a commercially PGM. There was a good linear relationship between PGM signal and concentration of E. coli or S. aureus as the bacteria model with high sensitivity. And our proposed biosensor was proved to be a rapid and reliable method for antimicrobial susceptibility testing within 4 h with consistent results of Minimum Inhibitory Concentrations (MICs) testing, providing a portable and convenient method to treat infected patients with correct antibiotics and reduce the production of antibiotic-resistant bacteria, especially for resource-limiting settings.

We present the use of DNA and peptide nucleic acid (PNA) molecular beacons (MBs) as sensitive indicators in microfluidic bioMEM devices. DNA and PNA MBs can be used to quantitatively study hybridization kinetics in real time in a polydimethyl siloxane (PDMS) microfluidic device. PNA MBs perform better than DNA MBs for the study of hybridization kinetics of rRNA targets in real time in microfluidic channels. We also demonstrate the use of PNA MBs for fast detection of bacterial cells in microfluidic channels. Using PNA MBs as detection probes will enable us to develop an integrated biosensor for the rapid and on-site detection and quantification of microbial pathogens in environmental and clinical samples.

Rapid and sensitive pathogen detection methods are critical for disease diagnosis and treatment. RPA-CRISPR/Cas12 systems have displayed remarkable potential in pathogen detection. A self-priming digital PCR chip is a powerful and attractive tool for nucleic detection. However, the application of the RPA-CRISPR/Cas12 system to the self-priming chip still has great challenges due to the problems of protein adsorption and two-step detection mode of RPA-CRISPR/Cas12. In this study, an adsorption-free self-priming digital chip was developed and a direct digital dual-crRNAs (3D) assay was established based on the chip for ultrasensitive detection of pathogens. This 3D assay combined the advantages of rapid amplification of RPA, specific cleavage of Cas12a, accurate quantification of digital PCR, and point-of-care testing (POCT) of microfluidics, enabling accurate and reliable digital absolute quantification of Salmonella in POCT. Our method can provide a good linear relationship of Salmonella detection in the range from 2.58 × 101 to 2.58 × 104 cells/mL with a limit of detection ∼0.2 cells/mL within 30 min in a digital chip by targeting the invA gene of Salmonella. Moreover, the assay could directly detect Salmonella in milk without nucleic acid extraction. Therefore, the 3D assay has the significant potential to provide accurate and rapid pathogen detection in POCT. This study provides a powerful nucleic detection platform and facilitates the application of CRISPR/Cas-assisted detection and microfluidic chips.

Advances in nucleic acid-based diagnostics of bacterial infections

A smart microfluidic platform for rapid multiplexed detection of foodborne pathogens

Rapid isolation and detection of aquaculture pathogens in an integrated microfluidic system using loop-mediated isothermal amplification

Rapid pathogen detection with bacterial-assembled magnetic mesoporous silica

We present a novel microfluidic platform for rapid detection and antimicrobial susceptibility assessment of bacteria directly from urine samples. Towards this end, we have developed a rapid and specific pathogen detection assay based on probe hybridization to bacterial ribosomal RNA. Our assay is implemented in an integrated microfluidic droplet platform capable of single-bacteria detection. The enhancement in speed and sensitivity enabled by the combination of our assay and platform facilitates rapid assessment of antimicrobial susceptibility directly from urine samples. Our platform can detect the effect of antibiotics on bacterial growth after as little as 10 min. drug exposure. We demonstrate the rapid response of our platform with 2 different classes of antibiotics.

Infections acquired from ingesting contaminated food and water poses an adverse effect on public health and safety, thus affecting nations’ economy. Technical approaches developed over years have contributed adequately to microbial detection in food and water, yet, unveiling spaces for more improvement on early and rapid detection of pathogens. This review highlights different strategy assessing bio-functionalized inorganic nanoparticles towards the detection of pathogens in food and water samples. Conjugates of several bio-receptors and inorganic nanoparticles showed rapid, real-time, repeatability, and appreciable limit of detection in targeted pathogens. A patent referenced in this study established the biocompatibility of bio-functionalized inorganic nanoparticles mechanism. Unique attributes exhibited by bio-functionalized inorganic nanoparticles showed potential and improvement of the existing bio-sensing pathogen detection methods. Each of the identified strategies described showed a promising pathway accommodating the development of simple, and even the fabrication of low-cost materials for easy detection of bacterial pathogens in food and water products.

Rapid, sensitive, and selective detection of live pathogens remains a key priority for quality control and risk assessment. While conventional methods often require complicated workflows, costly reagents, lab equipment, and are time‐consuming, rendering them inadequate for field testing and low‐resource settings. Increased attention has been drawn to developing alternative low‐cost and rapid methods to detect on‐site live pathogens in different environmental matrices. Among them, microfluidic devices that integrate various laboratory functions in a miniaturized manner have proven to be a promising tool for the rapid and sensitive detection of pathogens. Herein, the development of microfluidic devices specifically designed for the detection of live pathogens is discussed along a concise summary of novel microfluidics systems recently developed, contrasted to conventional methods regarding assay time, the limit of detection, and target organisms. These include a variety of micro total analysis systems (µTAS) and microfluidic paper‐based analytical devices (µPADs) in combination with molecular methods and traditional live cell detection techniques, such as cell culture, DNA intercalating dyes, resazurin, and immobilized bioreceptors (e.g., aptamers and capture antibodies). Furthermore, insights on the future perspectives of microfluidics for live pathogen detection with a highlight on the rapid and low‐cost method development for field testing are provided.

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

Rapid Detection of Airway Pathogens in Lung Transplant Donors

One-pot dual-CRISPR/Cas12b-mediated dUTP-LAMP for rapid and sensitive detection of foodborne pathogens without carryover contamination