Plasmonic Approach to Fluorescence Enhancement of Mesoporous Silica-Coated Gold Nanorods for Highly Sensitive Influenza A Virus Detection Using Lateral Flow Immunosensor.

Rapid diagnostic tests based on the lateral flow immunoassay (LFI) enable early identification of viral infection, owing to simple interpretation, short turnaround time, and timely isolation of patients to minimize viral transmission among communities. However, the LFI system requires improvement in the detection sensitivity to match the accuracy of nucleic acid amplification tests. Fluorescence-based LFIs are more sensitive and specific than absorption-based LFIs, but their performance is significantly affected by fundamental issues related to the quantum yield and photobleaching of fluorophores. Metal-enhanced fluorescence (MEF), which is a plasmonic effect in the vicinity of metallic nanoparticles, can be an effective strategy to improve the detection sensitivity of fluorescence-based LFIs. The key factors for obtaining a strong plasmonic effect include the distance and spectral overlap of the metal and fluorophore in the MEF system. In this study, MEF probes were designed based on core-shell nanostructures employing a gold nanorod core, mesoporous silica shell, and cyanine 5 fluorophore. To optimize the efficiency of MEF probes incorporated on the LFI platform (MEF-LFI), we experimentally and theoretically investigated the distance dependence of plasmonic coupling between cyanine 5 and gold nanorods by adjusting the shell thickness, resulting in significant fluorescence enhancement. The proposed MEF-LFI enabled highly sensitive detection of influenza A virus (IAV) nucleocapsid protein with a detection limit of 0.52 pg mL-1 within 20 min and showed high specificity and accuracy for determining IAV clinical samples. Overall, our findings demonstrate the potential of this method as an effective tool for molecular diagnosis under emergency conditions.

Adenosine Triphosphate Bioluminescence-Based Bacteria Detection Using Targeted Photothermal Lysis by Gold Nanorods.

Bacterial infections are common causes of morbidity and mortality worldwide; therefore, environmental contamination by bacterial pathogens represents a global public health concern. Consequently, a selective, rapid, sensitive, and in-field detection platform for detecting significant bacterial contamination is required to ensure hygiene and protect public health. Here, we developed a fast and simple platform for the selective and sensitive detection of bacteria by measuring adenosine triphosphate (ATP) bioluminescence following targeted photothermal lysis mediated by antibody-conjugated gold nanorods. This method employed both targeted photothermal lysis of bacteria by near-infrared (NIR) irradiation and highly selective detection of the lysed bacteria via ATP bioluminescence within 36 min (incubation, 30 min; NIR irradiation, 6 min). The use of the proposed method allowed limits of detection in pure solution of 12.7, 70.7, and 5.9 CFU for Escherichia coli O157:H7, Salmonella typhimurium, and Listeria monocytogenes, respectively. Additionally, bacteria were successfully detected on artificially inoculated plastic cutting boards. Furthermore, this method was highly specific, without cross-reaction among pathogenic bacteria. We believe that the proposed method has significant potential as an on-site diagnostic tool for applications associated with public health and environmental pollution monitoring.

Ultrasensitive Detection of Escherichia coli O157:H7 by Immunomagnetic Separation and Selective Filtration with Nitroblue Tetrazolium/5-Bromo-4-chloro-3-indolyl Phosphate Signal Amplification.

Here, we report an enhanced colorimetric method using enzymatic amplification with nitroblue tetrazolium (NBT)/5-bromo-4-chloro-3-indolyl phosphate (BCIP) precipitation for the ultrasensitive detection of Escherichia coli O157:H7 through immunomagnetic separation-selective filtration. Biotinylated anti- E. coli O157:H7 antibody and streptavidin-alkaline phosphatase were conjugated to the surface of magnetic nanoparticles, and E. coli O157:H7-conjugates complexes remained on the membrane filter surface. The resultant light brown spots on the membrane filter were amplified with NBT/BCIP solution to yield enzyme-catalyzed precipitation, which increased with an increasing E. coli O157:H7 concentration. E. coli O157:H7 was detected in pure samples with limits of detection of 10 and 6.998 colony-forming units (CFU)/mL through visual observation and measurement of optical density, respectively. The proposed method was applied to a lettuce sample inoculated with selective E. coli O157:H7, which was detected within 55 min without cross-reactivity to non-target bacteria. This enhanced colorimetric method has potential for on-site detection of food contaminants and environmental pollutants.

Homogeneous and selective detection of cadmium ions by forming fluorescent cadmium-protein nanoclusters.

We synthesized fluorescent Cd nanoclusters (CdNCs) through a protein-directed method, and the synthesis method was utilized for a homogeneous, ultrasensitive, and selective detection of cadmium ion (Cd2+). CdNCs were synthesized using a modified protein-directed method for developing a rapid Cd2+ detection system. For rapid Cd2+ detection, the reaction time was reduced by optimizing the reaction conditions such as temperature, reducing agent concentration, and protein concentration. The synthesized CdNCs had ca. 2 nm diameter and showed strong fluorescence at 485 nm under 365 nm UV light. The fluorescence of the CdNCs increased with increasing Cd2+ concentrations, and the limit of detection in deionized water was 15.68 fM. This method enables the detection of Cd2+ through the Cd concentration-dependent formation of fluorescent CdNCs in tap, fountain, and pond water samples with detection limits of 0.75, 7.65, and 48.2 fM, respectively. The sensitivity and specificity of our method are comparable to those of several existing methods for Cd2+ detection. Furthermore, the system enables the homogeneous detection of Cd2+ without separation and washing, thereby broadening its application in analytical chemistry.