Europium Nanoparticle-Based Lateral Flow Strip Biosensors Combined with Recombinase Polymerase Amplification for Simultaneous Detection of Five Zoonotic Foodborne Pathogens

Abstract

The five recognized zoonotic foodborne pathogens, namely, Listeria monocytogenes, Staphylococcus aureus, Streptococcus suis, Salmonella enterica and Escherichia coli O157:H7, pose a major threat to global health and social–economic development. These pathogenic bacteria can cause human and animal diseases through foodborne transmission and environmental contamination. Rapid and sensitive detection for pathogens is particularly important for the effective prevention of zoonotic infections. In this study, rapid and visual europium nanoparticle (EuNP)-based lateral flow strip biosensors (LFSBs) combined with recombinase polymerase amplification (RPA) were developed for the simultaneous quantitative detection of five foodborne pathogenic bacteria. Multiple T lines were designed in a single test strip for increasing the detection throughput. After optimizing the key parameters, the single-tube amplified reaction was completed within 15 min at 37 °C. The fluorescent strip reader recorded the intensity signals from the lateral flow strip and converted the data into a T/C value for quantification measurement. The sensitivity of the quintuple RPA-EuNP-LFSBs reached a level of 101 CFU/mL. It also exhibited good specificity and there was no cross-reaction with 20 non-target pathogens. In artificial contamination experiments, the recovery rate of the quintuple RPA-EuNP-LFSBs was 90.6–101.6%, and the results were consistent with those of the culture method. In summary, the ultrasensitive bacterial LFSBs described in this study have the potential for widespread application in resource-poor areas. The study also provides insights in respect to multiple detection in the field.

Breaking boundaries in microbiology: customizable nanoparticles transforming microbial detection

“…37 Different kinds of nanobiosensors, such as fabricated strips embedded with nanoparticles, nanocomposites decorated with spongins, multi-probe genomic biosensors, and nanocomposites-modified carbon screen-printed electrodes have already been shown to be extremely successful in bacterial cell detection. [38][39][40] A nanobiosensor termed the CRISPR/Cas12a-powered evanescent wave fluorescein Nanobiosensing platform (CREAT), was designed using the concept of multiple signal enhancement. The sensor has been validated and found to be far better than previous evanescent wave fluorescein nanobiosensors.…”

“…The samples taken for validating the sensor include chicken, pork, beef, lamb, duck, and milk. 39 In addition to identifying pathogens, it is crucial to develop sensors capable of detecting and isolating drug-resistant bacteria. To address this need, glycan-coated magnetic nanoparticles have been effectively deployed for the identification and detection of carbapenem-resistant E. coli in water and food.…”

“…37 Different kinds of nanobiosensors, such as fabricated strips embedded with nanoparticles, nanocomposites decorated with spongins, multi-probe genomic biosensors, and nanocomposites-modified carbon screen-printed electrodes have already been shown to be extremely successful in bacterial cell detection. 38–40…”

Nanomaterials-Integrated Electrochemical Biosensors as Pioneering Solutions for Zoonotic Disease Diagnosis

“…Jin and coworkers found out in a study focusing on the creation of lateral flow strip biosensors based on Eu nanoparticles combined with Recombinase Polymerase Amplification for concurrent identification of five food-borne infections. 88 To increase the detection capacity, many T lines were created to fit into a single test strip. Within 15 min at 37 °C, the single-tube amplified reaction was finished after the critical parameters were optimised.…”

Europium Nanoparticle-Based Lateral Flow Strip Biosensors for the Detection of Quinoxaline Antibiotics and Their Main Metabolites in Fish Feeds and Tissues

“…Lateral flow strip biosensors (LFSBs) are highly efficient biosensors and can be used to detect antibiotics in a rapid, simple, specific, sensitive, and reliable way. They also allow the visualization and quantification of target products [ 16 , 17 ].…”