Toothpick DNA extraction combined with handheld LAMP microfluidic platform for simple and rapid meat authentication.

Adulteration of meat is a global issue, necessitating rapid, inexpensive, and simple on-site testing methods. Therefore, the present study aimed to develop a one-minute toothpick-based DNA extraction method, a handheld microfluidic chip, and a smartphone-controlled portable analyzer for detecting multiple meat adulterations. A toothpick was inserted into the meat to promote DNA release and adsorption. Furthermore, a handheld microfluidic chip was designed for DNA elution on toothpicks and fluid distribution. Finally, a smartphone-actuated portable analyzer was developed to function as a heater, signal detector, and result reader. The portable device comprises a microcontroller, a fluorescence detection module, a step scanning unit, and a heating module. The proposed device is portable, and the app is user-friendly. This simple design, easy operation, and fast-response system could rapidly detect as little as 1% of simulated adulterated samples (following UK standards) within 40 min at a cost of less than USD 1 per test.

Ultra-sensitive fluorescent biosensor for multiple bacteria detection based on CDs/QDs@ZIF-8 and microfluidic fluidized bed.

An ultra-sensitive fluorescent biosensor based on CDs/QDs@ZIF-8 and microfluidic fluidized bed was developed for rapid and ultra-sensitive detection of multiple target bacteria. The zeolitic imidazolate frameworks (ZIF-8) act as the carrier to encapsulate three kinds of fluorescence signal molecules from the CDs/QDs@ZIF-8 signal amplification system. Besides, three kinds of target pathogenic bacteria were automatically, continuously, and circularly captured by the magnetic nanoparticles (MNPs) in the microfluidic fluidized bed. The neutral Na2EDTA solution was the first time reported to not only dissolve the ZIF-8 frameworks from the MNPs-bacteria-CDs/QDs@ZIF-8 sandwich complexes, but also release the CDs/QDs from sandwich complexes with no loss of fluorescence signal. Due to the advantages of signal amplification and automated sample pretreatment, the proposed fluorescent biosensor can simultaneously detect Escherichia coli O157:H7, Salmonella paratyphi A, and Salmonella paratyphi B as low as 101 CFU/mL within 1.5 h, respectively. The mean recovery in spiked milk samples can reach 99.18%, verifying the applicability of this biosensor in detecting multiple bacteria in real samples.

An instrument-free, integrated micro-platform for rapid and multiplexed detection of dairy adulteration in resource-limited environments.

In dairy industry, expensive yak's milk, camel's milk, and other specialty dairy products are often adulterated with low-cost cow's milk, goat's milk and so on. Currently, the detection of specialty dairy products typically requires laboratory settings and relies on skilled operators. Therefore, there is an urgent need to develop a multi-detection technology and on-site rapid detection technique to enhance the efficiency and accuracy of the detection of specialty dairy products. In this study, we introduced a fully integrated and portable microfluidic detection platform called Sector Self-Driving Microfluidics (SDM), designed to simultaneously detect eight common species-specific components in milk. SDM integrated nucleic acid extraction, purification, loop-mediated isothermal amplification (LAMP), and lateral flow strip (LFS) detection functions into a closed microfluidic system, enabling contamination-free visual detection. The SDM platform used a constant-temperature heating plate, powered by a mobile battery, eliminated the need for additional power support. The SDM platform achieved nucleic acid enrichment and transfer through magnetic force and liquid flow driven by capillary forces, operating without external pumps. The standalone SDM platform could detect dairy components with as low as 1% content within 1 h. Validation with 35 commercially available samples demonstrated 100% specificity and accuracy compared to the gold standard real-time PCR. The SDM platform provided the dairy industry with an efficient, convenient, and accurate detection tool, enabling rapid on-site testing at production facilities or sales points. This facilitated real-time monitoring of quality issues during the production process, quickly identifying potential risks and preventing substandard products from entering the market.

A smartphone-based diagnostic analyzer for point-of-care milk somatic cell counting.

BACKGROUND: Mastitis, a pervasive and detrimental disease in dairy farming, poses a significant challenge to the global dairy industry. Monitoring the milk somatic cell count (SCC) is vital for assessing the incidence of mastitis and the quality of raw cow's milk. However, existing SCC detection methods typically require large-scale instruments and specialized operators, limiting their application in resource-constrained settings such as dairy farms and small-scale labs. To address these limitations, this study introduces a novel, smartphone-based, on-site SCC testing method that leverages smartphone capabilities for milk somatic cell identification and enumeration, offering a portable and user-friendly testing platform. RESULTS: The central findings of our study demonstrate the effectiveness of the proposed method for counting milk somatic cells. Its on-site applicability, facilitated by the microfluidic chip, optical system, and smartphone integration, heralds a paradigm shift in point-of-care testing (POCT) for dairy farms and smaller laboratories. This approach bypasses complex processing and presents a user-friendly solution for real-time SCC monitoring in resource-limited settings. This device boasts several unique features: small size, low cost (<$1,000 total manufacturing cost and <$1 per test), and high accuracy. Remarkably, it delivers test results within just 2 min. Actual-sample testing confirmed its consistency with results from the commercial Bentley FTS/FCM cytometer, affirming the reliability of the proposed method. Overall, these results underscore the potential for transformative change in dairy farm management and laboratory testing practices. SIGNIFICANCE: In summary, this study concludes that the proposed smartphone-based method significantly contributes to the accessibility and ease of SCC testing in resource-limited environments. By fostering the use of POCT technology in food safety control, particularly in the dairy industry, this innovative approach has the potential to revolutionize the monitoring and management of mastitis, ultimately benefiting the global dairy sector.

Critical review and recent advances of emerging real-time and non-destructive strategies for meat spoilage monitoring.

Monitoring and evaluating food quality, especially meat quality, has received a growing interest to ensure human health and decrease waste of raw materials. Standard analytical approaches used for meat spoilage assessment suffer from time consumption, being labor-intensive, operation complexity, and destructiveness. To overcome shortfalls of these traditional methods and monitor spoilage microorganisms or related metabolites of meat products across the supply chain, emerging analysis devices/systems with higher sensitivity, better portability, on-line/in-line, non-destructive and cost-effective property are urgently needed. Herein, we first overview the basic concepts, causes, and critical monitoring indicators associated with meat spoilage. Then, the conventional detection methods for meat spoilage are outlined objectively in their strengths and weaknesses. In addition, we place the focus on the recent research advances of emerging non-destructive devices and systems for assessing meat spoilage. These novel strategies demonstrate their powerful potential in the real-time evaluation of meat spoilage.

Amplification-free quantitative detection of genomic DNA using lateral flow strips for milk authentication.

With the globalization and complexity of the food supply chain, the market is becoming increasingly competitive and food fraudulent activities are intensifying. The current state of food detection faced two primary challenges. Firstly, existing testing methods were predominantly laboratory-based, requiring complex procedures and precision instruments. Secondly, there was a lack of accurate and efficient quantitative detection methods. Taking cow's milk as an example, this study introduced a novel method for nucleic acid quantification in dairy products, based on lateral flow strips (LFS). The core idea of this method is to design single-stranded DNA (ssDNA) probes to hybridize with mitochondrial genes, which are abundant, stable, and species-specific in dairy products, as detection targets. Drawing inspiration from the principles of nucleic acid amplification, this research innovatively established a new DNA hybridization method, named LAMP-Like Hybridization (HybLAMP-Like). Leveraging the denaturation and DNA polymerization functions of the bst enzyme, efficient binding of the probe and template strand was achieved. This method eliminated the need for nucleic acid amplification, simplifying the procedure and mitigating aerosol contamination, thereby ensuring the accuracy of the detection results. The method exhibited exceptional sensitivity, capable of detecting extremely low to 12.5 ng in visual inspection and 3.125 ng when using a reader. In terms of practicality, it could achieve visual detection of cow's milk content as low as 1% in adulterated dairy products. When combined with a portable LFS reader, it also enabled precise quantitative analysis of milk adulteration.

A Person-Environment Fit Model to Explain Information and Communication Technologies-Enabled After-Hours Work-Related Interruptions in China.

Given the ubiquitous nature of mobile devices and information and communication technologies (ICT), after-hours work-related interruptions (AHWI) occur anywhere and anytime in China. In the current study, an alternative person-environment (P-E) fit model of ICT-enabled AHWI, hereafter referred to as IAWI, that treats polychronic variables as moderated solutions are presented. A cross-sectional survey among 277 Chinese employees (average age: 32.04 years) was conducted in September 2022 and tested by PLS-structural equation modeling to validate our hypotheses. The results indicated that IAWI had a positive influence on employees' innovative job performance and in-role job performance (ß = 0.139, p < 0.05; ß = 0.200, p < 0.01; ß = 0.298, p < 0.001). Moreover, among employees with higher levels of polychronicity, the heightened effects of IAWI on innovative job performance were increased (ß = 0.112, p < 0.05). This study offers implications for employees: under IAWI situations, they could search for a person-environment (P-E) that is fit to buffer the negative aspects of IAWI, consequently increasing their innovative job performance and in-role job performance. Future research could extend beyond this framework to explore employees' IAWI and job performance balance.

Integrating microneedle DNA extraction to hand-held microfluidic colorimetric LAMP chip system for meat adulteration detection.

Most microfluidic-based "sample-in-result-out" systems suffer sophisticated microfluidic production processes, high-cost chips, and expensive instruments. They cannot be used in the meat market as well as farmer's markets in rural areas. Here, we developed a hand-held microfluidic chip system for on-site meat species qualitative authentication detection which integrated a simple microneedle DNA extraction and a visual loop-mediated isothermal amplification (LAMP). The chip can be used by easily pricking meat samples, simply hand-shaking the chip, and readily available isothermal heating instead of a complicated DNA extraction process and microfluidic control device. The system demonstrates high specificity and sensitivity for selected six species of meat samples and low to 1% simulated adulteration could be detected within 60 min. Besides, the whole cost was less than 1 dollar. The integrated hand-held microfluidic detection system offers a simple, fast, low-cost "sample-in-result-out" point-of-care device which could be extended to medical diagnosis and animal/plant disease identification.

Biosensors Coupled with Signal Amplification Technology for the Detection of Pathogenic Bacteria: A Review.

Foodborne disease caused by foodborne pathogens is a very important issue in food safety. Therefore, the rapid screening and sensitive detection of foodborne pathogens is of great significance for ensuring food safety. At present, many research works have reported the application of biosensors and signal amplification technologies to achieve the rapid and sensitive detection of pathogenic bacteria. Thus, this review summarized the use of biosensors coupled with signal amplification technology for the detection of pathogenic bacteria, including (1) the development, concept, and principle of biosensors; (2) types of biosensors, such as electrochemical biosensors, optical biosensors, microfluidic biosensors, and so on; and (3) different kinds of signal amplification technologies applied in biosensors, such as enzyme catalysis, nucleic acid chain reaction, biotin-streptavidin, click chemistry, cascade reaction, nanomaterials, and so on. In addition, the challenges and future trends for pathogenic bacteria based on biosensor and signal amplification technology were also discussed and summarized.

Multicolor Coding Up-Conversion Nanoplatform for Rapid Screening of Multiple Foodborne Pathogens.

Technologies for rapid screening of multiple foodborne pathogens have been urgently needed because of the complex food matrix and high outbreaks of foodborne diseases. In this study, multicolor coding up-conversion nanoparticles (UCNPs) were synthesized and applied for rapid and simultaneous detection of five kinds of foodborne pathogens. The multicolor coding UCNPs were obtained through doping different concentrations of a sensitizer (Yb3+) on the shell of the synthesized NaYF4:Yb3+, Tm3+ (20%/2%)@NaYF4:Yb3+, and Er3+ (x %/2%) core/shell nanocrystals. All the UCNPs could emit red and green luminescence simultaneously once excited with near-infrared wavelength (980 nm), and the ratio of red and green (R/G ratio) emission light intensity of each kind of UCNPs varied depending on the Yb3+ doping concentration. In addition, the magnetic nanoparticles (MNPs) modified with the monoclonal antibodies (mAbs) against the target bacteria were used to capture and separate the bacteria, resulting in obtaining the MNP-bacterium complexes. Different UCNPs with multicolor coding acted as signal probes were also modified with the mAbs to react with the MNP-bacterium complexes to form the MNP-bacterium-UCNP sandwich complexes. After the sandwich complexes were excited with a wavelength of 980 nm, the obtained R/G ratios and the green photoluminescence intensity (PL intensity) could be used to distinguish and quantitatively detect foodborne pathogens, respectively. This proposed nanoplatform could detect five foodborne pathogens simultaneously within 2 h with good sensitivity and specificity, showing great potential for multiplex detection of other targets in the fields of medical diagnosis and food security.

A microfluidic biosensor for rapid and automatic detection of Salmonella using metal-organic framework and Raspberry Pi.

Rapid screening of pathogenic bacteria contaminated foods is crucial to prevent food poisoning. However, available methods for bacterial detection are still not ready for in-field screening because culture is time-consuming; PCR requires complex DNA extraction and ELISA lacks sensitivity. In this study, a microfluidic biosensor was developed for rapid, sensitive and automatic detection of Salmonella using metal-organic framework (MOF) NH2-MIL-101(Fe) with mimic peroxidase activity to amplify biological signal and Raspberry Pi with self-developed App to analyze color image. First, the target bacteria were separated and concentrated with the immune magnetic nanobeads (MNBs), and labeled with the immune MOFs to form MNB-Salmonella-MOF complexes. Then, the complexes were used to catalyze colorless o-phenylenediamine and H2O2 to produce yellow 2,3-diaminophenazine (DAP). Finally, the image of the catalysate was collected under the narrow-band blue light and analyzed using the Raspberry Pi App to determine the bacterial concentration. The experimental results showed that this biosensor was able to detect Salmonella Typhimurium from 1.5 × 101 to 1.5 × 107 CFU/mL in 1 h with the lower detection limit of 14 CFU/mL. The mean recovery for Salmonella in spiked chicken meats was ~112%. This biosensor integrating mixing, separation, labelling and detection onto a single microfluidic chip has demonstrated the merits of automatic operation, fast reaction, less reagent and small size, and is promising for in-field detection of foodborne bacteria.

An ultrasensitive impedance biosensor for Salmonella detection based on rotating high gradient magnetic separation and cascade reaction signal amplification.

An impedance biosensor using rotary magnetic separation and cascade reaction was developed for rapid and ultrasensitive detection of Salmonella typhimurium. First, magnetic nanoparticles (MNPs) modified with anti-Salmonella monoclonal antibodies were injected into a capillary at the presence of a rotary high gradient magnetic field, which was rotated by a stepper motor. Then, a bacterial sample was injected into the capillary and the target bacteria were continuous-flow captured onto the MNPs. After organic-inorganic hybrid nanoflowers were prepared using manganese dioxide (MnO2), glucose oxidase (GOx) and anti-Salmonella polyclonal antibodies (pAbs), they were injected to label the bacteria, resulting in the formation of MNP-bacteria-nanoflower sandwich complexes. Finally, glucose (low conductivity) was injected and oxidized by GOx on the complexes to produce H2O2 (low conductivity) and gluconic acid (high conductivity), leading to impedance decrease. Besides, the produced H2O2 triggered a cascade reduction of MnO2 into Mn2+, leading to further impedance decrease. The impedance changes were measured using an interdigitated microelectrode and used to determine the concentration of target bacteria. This biosensor was able to detect Salmonella ranging from 101 to 106 CFU/mL in 2 h with a low detection limit of 101 CFU/mL and a mean recovery of 100.1% for the spiked chicken samples.

An impedance biosensor based on magnetic nanobead net and MnO2 nanoflowers for rapid and sensitive detection of foodborne bacteria.

Screening of pathogenic bacteria in foods is an effective way to prevent foodborne diseases. In this study, an impedance biosensor was developed for rapid and sensitive detection of Salmonella typhimurium using multiple magnetic nanobead (MNB) nets in a ring channel for continuous-flow separation of target bacteria from 10 mL of sample, manganese dioxide nanoflowers (MnO2 NFs) for efficient amplification of biological signal, and an interdigitated microelectrode for sensitive measurement of impedance change. First, the MNBs modified with capture antibodies were vortically injected from outer periphery of this ring channel to form multiple ring MNB nets at specific locations with high gradient magnetic fields. Then, the bacterial sample was continuous-flow injected, resulting in specific capture of target bacteria onto the nets, and the MnO2 NFs modified with detection antibodies were injected to form MNB-bacteria-MnO2 NF complexes. After the complexes were washed with deionized water to remove excessive nanoflowers and residual ions, H2O2 with poor conductivity was injected to reduce MnO2 NFs to conductive Mn2+ at neutral medium, leading to impedance decrease. Finally, impedance change was measured using the microelectrode for quantitative determination of Salmonella. This biosensor was able to separate ~60% of Salmonella from 10 mL of bacterial sample and detect Salmonella with a linear range of 3.0 × 101 to 3.0 × 106 CFU/mL in 1.5 h with lower detection limit of 19 CFU/mL. This biosensor might be further improved with higher sensitivity using a larger volume (100 mL or more) for routine screening of foodborne bacteria without bacterial pre-culture.

A sensitive immunoassay for simultaneous detection of foodborne pathogens using MnO2 nanoflowers-assisted loading and release of quantum dots.

In this study, a sensitive immunoassay using immunomagnetic nanobeads (MNBs), manganese dioxide nanoflowers (MnO2 NFs) and quantum dots (QDs) was developed for simultaneous detection of E. coli O157: H7 and Salmonella typhimurium. MnO2 NFs were synthesized, functionalized and incubated with QDs to obtain QDs@MnO2 nanocomposites, followed by modification with antibodies (pAbs) to obtain pAb-QDs@MnO2 nanocomposites (QM NCs). Target bacteria were first conjugated with MNBs and QM NCs to form MNB-bacteria-QM complexes. Then, QDs were quickly released from the complexes using glutathione to reduce MnO2 to Mn2+. Finally, fluorescent intensity at characteristic wavelength was measured by optical detector to determine target bacteria. This immunoassay could simultaneously and quantitatively detect E. coli from 1.5 × 101 to 1.5 × 106 CFU/mL with detection limit of 15 CFU/mL and Salmonella from 4.0 × 101 to 4.0 × 106 CFU/mL with detection limit of 40 CFU/mL in 2 h. The mean recovery for both bacteria in spiked chicken samples was ∼96%.

A Microfluidic Biosensor Based on Magnetic Nanoparticle Separation, Quantum Dots Labeling and MnO2 Nanoflower Amplification for Rapid and Sensitive Detection of Salmonella Typhimurium.

Screening of foodborne pathogens is an effective way to prevent microbial food poisoning. A microfluidic biosensor was developed for rapid and sensitive detection of Salmonella Typhimurium using quantum dots (QDs) as fluorescent probes for sensor readout and manganese dioxide nanoflowers (MnO2 NFs) and as QDs nanocarriers for signal amplification. Prior to testing, amino-modified MnO2 nanoflowers (MnO2-NH2 NFs) were conjugated with carboxyl-modified QDs through EDC/NHSS method to form MnO2-QD NFs, and MnO2-QD NFs were functionalized with polyclonal antibodies (pAbs) to form MnO2-QD-pAb NFs. First, the mixture of target Salmonella Typhimurium cells and magnetic nanoparticles (MNPs) modified with monoclonal antibodies (mAbs) was injected with MnO2-QD-pAb NFs into a microfluidic chip to form MNP-bacteria-QD-MnO2 complexes. Then, glutathione (GSH) was injected to dissolve MnO2 on the complexes into Mn2+, resulting in the release of QDs. Finally, fluorescent intensity of the released QDs was measured using the fluorescent detector to determine the amount of Salmonella. A linear relationship between fluorescent intensity and bacterial concentration from 1.0 × 102 to 1.0 × 107 CFU/mL was found with a low detection limit of 43 CFU/mL and mean recovery of 99.7% for Salmonella in spiked chicken meats, indicating the feasibility of this biosensor for practical applications.

A colorimetric immunosensor for determination of foodborne bacteria using rotating immunomagnetic separation, gold nanorod indication, and click chemistry amplification.

A colorimetric immunosensor was developed for the determination of Salmonella Typhimurium using rotating magnetic separation, gold nanorod (GNR) indication, and click chemistry amplification. The target bacteria were first separated from large-volume sample using a rotating magnetic field and a small amount (50 µg) of immunomagnetic nanoparticles (MNPs), resulting in the forming of magnetic bacteria. Then, the magnetic bacteria were conjugated with catalase (CAT)-labeled antibodies, which were synthesized using trans-cyclooctene/1,2,4,5-tetrazine click chemistry reaction, resulting in the forming of enzymatic bacteria. Then the CATs on the enzymatic bacteria were used to decompose an excessive amount of hydrogen peroxide (H2O2), the remaining H2O2 was mixed with horseradish peroxidase to etch the GNRs, resulting in color change and absorbance peak shift of the GNRs. Finally, the peak shift was measured and analyzed for the quantitative determination of target bacteria. This immunosensor was able to detect Salmonella Typhimurium with a linear range of 101-105 CFU mL-1 in 3 h with a low detection limit of 35 CFU mL-1. The mean recovery for Salmonella Typhimurium in spiked chicken samples was 109%. Graphical abstractSchematic representation of a colorimetric immunosensor for the determination of Salmonella Typhimurium as low as 35 CFU mL-1 using rotating magnetic separation of Salmonella from a large-volume sample, click chemistry reaction of catalase with antibodies for signal amplification, and HRP-mediated gold nanorod etching for result indication.

Panning anti-LPS nanobody as a capture target to enrich Vibrio fluvialis.

Vibrio fluvialis is considered as a human pathogen in developing countries. This bacterium is widely distributed in seawater and harbors that contains traces of salt. V. fluvialis can cause human enteritis and diarrhea, which has broken out at a global scale. Lipopolysaccharide (LPS) is a key bacterial antigen used to classify V. fluvialis serogroups. In this research, phage display technology was adopted to isolate nanobodies from a naïve phage library by using LPS as the target antigen. The isolated nanobody was tested in LPS ELISA and bacterial enzyme-linked immunosorbent assay Nanobody V23 had a high affinity toward the pathogen and was utilized to synthesize immunomagnetic beads for the enrichment of V. fluvialis. The capture efficiency of the immunomagnetic beads against V. fluvialis was 90.7 ±â€¯3.2% (N = 3) through the plate-counting method. We generated a high-affinity nanobody against LPS from V. fluvialis and developed a rapid method of enriching V. fluvialis by using immunomagnetic beads.

A capillary biosensor for rapid detection of Salmonella using Fe-nanocluster amplification and smart phone imaging.

Early screening of foodborne pathogenic bacteria is a key to prevent and control foodborne diseases. This study intended to develop a capillary biosensor for rapid and sensitive detection of Salmonella using the multi-column capillary for easy operation, the Fe-nanoclusters (FNCs) for signal amplification and the smart phone APP for image analysis. The multi-column capillary was successively preloaded the magnetic nanoparticle (MNP) column, the FNC column, two phosphate buffer solution with 0.05% Tween 20 (PBST) columns and the HCl column and the columns were separated by air gap. The iron spiral mixer was fabricated to accelerate the mixing, and the multi-ring magnets was developed to transfer the MNPs and their conjugates from column to column. The target bacteria were captured by the MNPs to form the magnetic bacteria and then conjugated with the FNCs to form the nanocluster bacteria. After washing with PBST, the nanocluster bacteria were transferred into the HCl column, and the iron ions were released and reacted with potassium hexacyanoferrate to form the Prussian Blue, which was finally measured and analyzed using the Hue-Saturation-Lightness color space based smartphone APP for the determination of the target bacteria. This proposed biosensor exhibited a wide linear range for detection of Salmonella typhimurium with the lower detection limit of 14 CFU/mL. The mean recovery of the target bacteria in spiked chicken samples was ~105.0%, indicating the applicability of this biosensor. The proposed biosensor had the potential for in-field detection of foodborne pathogens.

Rapid and sensitive detection of Escherichia coli O157:H7 using coaxial channel-based DNA extraction and microfluidic PCR.

In this study, a rapid and sensitive method for detection of Escherichia coli O157:H7 using the coaxial channel-based DNA extraction and the microfluidic PCR was proposed and verified. The magnetic silica beads were first pumped into the coaxial channel, which was captured in the coaxial channel more uniformly by applying the multiring high-gradient magnetic field. After the E. coli O157:H7 cells were lysed with the lysis buffer to release the DNA, the improved coaxial channel was used to efficiently extract the DNA, followed by washing with ethanol to remove the residual proteins and eluting with a small volume of deionized water to obtain the purified and concentrated DNA. Finally, the obtained DNA was amplified and determined using the microfluidic PCR. This proposed bacteria detection method was able to detect E. coli O157:H7 as low as 12 cfu/mL when the large volume (10 mL) of bacterial sample was used, and the recovery of E. coli O157:H7 in the spiked milk samples ranged from 97.4 to 100.6%. This proposed bacteria detection method has shown great potential to detect lower concentration of E. coli O157:H7 from larger volumes of sample.

A sensitive biosensor using double-layer capillary based immunomagnetic separation and invertase-nanocluster based signal amplification for rapid detection of foodborne pathogen.

Combining double-layer capillary based high gradient immunomagnetic separation, invertase-nanocluster based signal amplification and glucose meter based signal detection, a novel biosensor was developed for sensitive and rapid detection of E. coli O157:H7 in this study. The streptavidin modified magnetic nanobeads (MNBs) were conjugated with the biotinylated polyclonal antibodies against E. coli O157:H7 to form the immune MNBs, which were captured by the high gradient magnetic field in the double-layer capillary to specifically separate and efficiently concentrate the target bacteria. Calcium chloride was used with the monoclonal antibodies against E. coli O157:H7 and the invertase to form the immune invertase-nanoclusters (INCs), which were used to react with the target bacteria to form the MNB-bacteria-INC complexes in the capillary. The sucrose was then injected into the capillary and catalyzed by the invertase on the complexes into the glucose, which was detected using the glucose meter to obtain the concentration of the glucose for final determination of the E. coli O157:H7 cells in the sample. A linear relationship between the readout of the glucose meter and the concentration of the E. coli O157:H7 cells (from 102 to 107 CFU/mL) was found and the lower detection limit of this biosensor was 79 CFU/mL. This biosensor might be extended for the detection of other foodborne pathogens by changing the antibodies and has shown the potential for the detection of foodborne pathogens in a large volume of sample to further increase the sensitivity.