Increasing foodborne illnesses have led to global health and economic burdens. E. coli O157:H7 is one of the most common disease-provoking pathogens and known to be lethal Shiga toxin-producing E. coli (STEC) strains. With a low infection dose in addition to person-to-person transmission, STEC infections are easily spread. As a result, specific and rapid testing methods to identify foodborne pathogens are urgently needed. Nanozymes have emerged as enzyme-mimetic nanoparticles, demonstrating intrinsic catalytic activity that could allow for rapid, specific, and accurate pathogen identification in the agrifood industry. In this study, we developed a sensitive nanoplatform based on the traditional ELISA assay with the synergistic properties of gold and iron oxide nanozymes, replacing the conventional enzyme horseradish peroxidase (HRP). We designed an easily interchangeable sandwich ELISA composed of a novel, multifunctional magneto-plasmonic nanosensor (MPnS) with target antibodies (MPnS-Ab). Our experiments demonstrate a 100-fold increase in catalytic activity in comparison to HRP with observable color changes within 15 min. Results further indicate that the MPnS-Ab is highly specific for E. coli O157:H7. Additionally, effective translatability of catalytic activity of the MPnS technology in the lateral flow assay (LFA) platform is also demonstrated for E. coli O157:H7 detection. As nanozymes display more stability, tunable activity, and multi-functionality than natural enzymes, our platform could provide customizable, low-cost assay that combines high specificity with rapid detection for a variety of pathogens in a point-of-care setup.
Detection of bacterial contaminants in blood and platelet concentrates (PCs) continues to be challenging in clinical settings despite available current testing methods. At the same time, it is important to detect the low bacterial contaminants present at the time of transfusion. Herein, we report the design and synthesis of a dual-modal magnetofluorescent nanosensor (MFnS) by integrating magnetic relaxation and fluorescence modalities for the wide-range detection of blood-borne pathogens. In this study, functional MFnSs are designed to specifically detect Staphylococcus epidermidis and Escherichia coli, two of the predominant bacterial contaminants of PCs. Specific interaction between the target pathogen and functional MFnS resulted in the change of water proton's magnetic relaxation time, indicative of sensitive detection of the target bacteria from low to high colony-forming units (CFUs). In addition, the acquired magnetic relaxation signal of the MFnS further facilitated quantitative assessment of the slow and fast growth kinetics of target pathogens. Moreover, the presence of fluorescence modality in MFnS allowed for the detection of multicontaminants. Bacterial detection was also performed in complex media including whole blood and PCs, which further demonstrated its robust detection sensitivity. Overall, our study indicated that the designer MFnS will have potential for the wide-range detection of blood-borne pathogens and features desirable qualities including timeliness, sensitivity, and specificity.
Infection with the Zika virus (ZIKV) is an ongoing problem especially as accurate, cost-effective testing remains unresolved. In addition, coinfection occurs with both the Dengue virus (DENV) and ZIKV which leads to cross-reactivity between the flaviviruses and can result in false positives and inaccurate testing. This supports the current need for a simple assay that can detect Zika antibodies sensitively that at the same time can differentiate between cross-reactive antibodies. In this study, we developed customizable magnetic relaxation nanosensors (MRnS) conjugated to various ligands, which included ZIKV (ZENV, zika domain III and NS1) and DENV proteins for specific detection of cross-reactive Zika and Dengue antibodies. Binding interactions between functional MRnS and corresponding targets resulted in the change in spin−spin magnetic relaxation time (T2MR) of water protons, allowing for a rapid and simple means by which these interactions were detected and quantified. Our results show the detection of Zika antibodies within minutes at concentrations as low as 20 nM and display high specificity, reproducibility, and analytical sensitivity. Furthermore, a mixture of functional MRnS was used for the one-step simultaneous detection and differentiation of Zika and Dengue infections. These results demonstrate high specificity and sensitivity for the detection of ZIKV and DENV despite coinfections in both simple and complex media. Overall, our magnetic nanoplatform could be used as a rapid and sensitive assay for the detection of not only Zika-and Dengue-related testing but can be further applied to serological samples of any other pathogens.
Frequent outbreaks of food-borne pathogens, particularly E. coli O157:H7, continue to impact human health and the agricultural economy tremendously. The required cell count for this pathogenic strain of E. coli O157:H7 is relatively low and hence it is vital to detect at low colony forming unit (CFU) counts. Available detection methods, though sensitive, fall short in terms of timeliness and often require extensive sample processing. To overcome these limitations, we propose a novel magneto-plasmonic nanosensor (MPnS) by integrating surface plasmon resonance (SPR) properties with spin–spin magnetic relaxation (T2 MR) technology. We engineered MPnS by encapsulating several gold nanoparticles (GNPs) within the polymer-coating of iron oxide nanoparticles (IONPs). First, the polyacrylic acid (PAA)-coated IONPs were synthesized using a solvent precipitation method, then gold chloride solution was used to synthesize GNPs and encapsulate them within the PAA-coatings of IONPs in one step. A magnetic separation technique was used to purify the MPnS and the presence of GNPs within IONPs was characterized using transmission electron microscopy (TEM), energy dispersive x-ray spectroscopy (EDS), and other spectroscopic methods. The synthesized MPnS exhibits MR relaxation properties while possessing amplified optical properties than conventional GNPs. This allows for rapid and ultrasensitive detection of E. coli O157:H7 by SPR, T2 MR, and colorimetric readout. Experiments conducted in simple buffer and in milk as a complex media demonstrated that our MPnS-based assay could detect as low as 10 CFUs of this pathogenic strain of E. coli O157:H7 in minutes with no cross-reactivity. Overall, the formulated MPnS is robust and holds great potential for the ultrasensitive detection of E. coli O157:H7 in a simple and timely fashion. Moreover, this platform is highly customizable and can be used for the detection of other foodborne pathogens.
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