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The need for an accurate and rapid diagnosis platform for highly contagious viruses, such as pandemic viruses, has rapidly increased. Conventional diagnostic methods in the initial stage of quarantine for the prevention of epidemic spread have encountered limitations such as a long operation time, occupation of a large space, and high cost per single operation. Here, a highly sensitive and selective portable diagnostic platform based on solid‐phase photonic polymerase chain reaction (SP‐PCR) is reported. The thermal cycle is operated by the photothermal energy converted from the photon energy of a light‐emitting diode (LED) through a gold layer to phonons. The formation of an amplicon, which is immobilized through N‐heterocyclic carbene, is confirmed through fluorescence on the surface of the miniaturized SP‐PCR chip. The practical applicability of the newly developed diagnosis platform for the recognition of the presence of a SARS‐CoV‐2‐specific genome sequence is demonstrated utilizing 100 clinical samples, including 72 COVID‐19‐positive clinical samples and 28 negative samples. [ABSTRACT FROM AUTHOR]

Interfacial chemicals for metal surface functionalization were developed for applications of high water dispersibility and environmental stability. Metal nanomaterials, i.e., gold nanoparticles (AuNPs), were synthesized by introducing various interfacial chemicals, to improve the hydrophilicity of biosensors, such as those used in fluorescence resonance energy transfer (FRET) and lateral flow assay (LFA), respectively. Previously, thiolated AuNPs (SH-AuNPs) exhibited colloidal instability by forming irreversible aggregates in extreme environmental conditions; this phenomenon led to limitations such as poor sensitivity and reproducibility, in terms of biosensor application fields. Therefore, the development of novel interfacial chemicals remained a challenge for AuNP-based biosensor applications. Here, we first synthesized and demonstrated an ultra-stable AuNP functionalization by introducing N-heterocyclic carbene (NHC) compounds with a polyethylene glycol chain and azide terminal groups (NHC-AuNPs). The high binding energy of NHC-AuNPs compared with SH-AuNPs was demonstrated by density functional theory simulation, with NHC-AuNPs showing an unprecedented stability in extreme environmental conditions with varying ranges of pH, salts, and temperature; in particular, ultra-stability was observed in condition by freezing/thawing over 120 times. NHC-AuNPs were applied FRET and LFA biosensors and showed excellent sensing performances. Based on the results, NHC-AuNPs can be introduced for performance improvement in the development of diagnostic platforms to utilize in extreme environmental conditions. [ABSTRACT FROM AUTHOR]

The development of a rapid, simple, inexpensive, and sensitive nucleic acid assay is critical in ultrafast diagnostic systems to prevent the rapid spread of novel infectious agents. However, current representative diagnostic methods, such as the polymerase chain reaction (PCR), are bulky, expensive, complex, and time-consuming. The plasmonic photothermal effect of plasmonic materials is widely used in biological applications because of excellent properties, such as strong optical properties and enhanced local electromagnetic field. PCR methods using plasmonic nanostructures are used to overcome the limitations of conventional PCR. Therefore, plasmonic photothermal-based PCR exhibit a rapid ramping rate, ultrafast amplification, high sensitivity, cost-effectiveness, and compactness, which can be used as a point-of-care device in situ. This study reviewed the mechanisms of plasmonic PCR and various materials, such as organic, inorganic, and hybrid materials, used in PCR. Furthermore, the properties, light source, amplification efficiency, and monitoring method of classified plasmonic materials were compared. This review introduced the disease diagnosis application using plasmonic PCR and evaluated the prospect of potential future applications as next-generation PCR. [ABSTRACT FROM AUTHOR]

Fatal pathogenic organisms have been detected by various point‐of‐care tests (PoCTs) in clinical samples. PoCTs based on portable electrical devices are able to detect organisms in simple and rapid method. Although electrical PoCT devices show ultra sensitivity when detecting pathogens in standard samples, their industrialization has been limited due to low reproducibility, sensitivity, and specificity, resulting from nonspecific binding issues by media. In this research, a perpendicular N‐heterocyclic carbene self‐assembled monolayer (NHC‐SAM) conjugated on graphene micropattern field‐effect transistors (NGMFETs) is first demonstrated for monitoring pathogens. Two single bonds of NHCSAM on GM are investigated via density functional theory (DFT) and surface analyses. Bioreceptors are conjugated on the side gate‐modulated NGMFET for ultra‐stable on‐site detection. The side‐gated BNGMFETs system displayed remarkable performance in the detection of the following: i) SARS‐CoV‐2 spike protein S1 (limit of detection (LOD): ≈10 pg mL−1) in CTM, ii) O. tsutsugamushi Ab (LOD: ≈1 pg mL−1), and iii) E. coli (LOD: 10° CFU mL−1) in serum. Compared to commercial methodology, the platform presents 102 times higher LODs towards those pathogens in clinical samples with higher reproducibility. Based on these results, side‐gated BNGMFETs can be utilized as universal PoCT during pandemics. [ABSTRACT FROM AUTHOR]

Bitterness elicits unpleasant sensations in humans, which can hinder the acceptance of foods and medication adherence. Therefore, identifying and masking bitter tastes is crucial for developing palatable foods and promoting medication compliance in the food and pharmaceutical industries. To achieve this, employing agonism and antagonism of bitter taste receptors as effective strategies at the molecular level is essential. In this study, a bioelectronic tongue is developed to characterize the agonism and antagonism of bitter taste receptors. The human bitter taste receptors hTAS2R16 and hTAS2R31 are produced using an Escherichia coli expression system and reconstituted into nanodiscs (NDs). Subsequently, hTAS2R16‐ and hTAS2R31‐NDs are immobilized on the surface of graphene field‐effect transistors (FETs) to construct bioelectronic tongues. The developed system sensitively detected the bitter agonists, salicin and saccharin, at concentrations as low as 100 fM, with high selectivity in real‐time. The dose‐dependent curves shifted and K values decreased by the antagonists of hTAS2R16 and hTAS2R31, indicating antagonism‐based masking of bitter taste. Therefore, the developed bioelectronic tongue holds promise for identifying bitter tastes and evaluating the masking of bitterness based on the agonism and antagonism of hTAS2Rs. [ABSTRACT FROM AUTHOR]

A molecular diagnosis of the respiratory syncytial virus (RSV) without bulky and expensive instrumentation is of great importance for the early detection and prevention in a fast‐spreading pandemic. However, the current representative diagnostic methods have the limitation of being time‐consuming, cost, the processing time for polymerase chain reaction (PCR), and inaccurate for lateral flow assay (LFA), representatively. Herein, an integrated photonic digital PCR (dPCR) is developed with high‐velocity photonic scanner for in situ fluorescence detection by introducing the N‐heterocyclic carbene self‐assembled monolayer‐based Au film to prevent the quenching effect. The on‐site rapid molecular diagnostic platform shows the driving of 40 cycles in under 8 min and fluorescence scanning in under 7 min, resulting in a total analysis time within 15 min. In particular, the technology clearly demonstrates the classification of SARS‐CoV‐2 patients and healthy controls (99% in sensitivity, 98.6% in specificity, and 96.4% in accuracy with RdRp gene), comparing with standard RT‐qPCR. This platform can be utilized for prompt point‐of‐care molecular diagnostics in early diagnosis and large‐scale prevention of next pandemic spreading for upcoming infectious diseases and for the distinction diagnosis with other RSV. [ABSTRACT FROM AUTHOR]

Herein, we developed a bio-functionalized solution-immersed silicon (SIS) sensor at the single-cell level to identify Erwinia amylovora (E. amylovora), a highly infectious bacterial pathogen responsible for fire blight, which is notorious for its rapid spread and destructive impact on apple and pear orchards. This method allows for ultra-sensitive measurements without pre-amplification or labeling compared to conventional methods. To detect a single cell of E. amylovora, we used Lipopolysaccharide Transporter E (LptE), which is involved in the assembly of lipopolysaccharide (LPS) at the surface of the outer membrane of E. amylovora, as a capture agent. We confirmed that LptE interacts with E. amylovora via LPS through in-house ELISA analysis, then used it to construct the sensor chip by immobilizing the capture molecule on the sensor surface modified with 3′-Aminopropyl triethoxysilane (APTES) and glutaraldehyde (GA). The LptE-based SIS sensor exhibited the sensitive and specific detection of the target bacterial cell in real time. The dose–response curve shows a linearity (R2 > 0.992) with wide dynamic ranges from 1 to 107 cells/mL for the target bacterial pathogen. The sensor showed the value change (dΨ) of approximately 0.008° for growing overlayer thickness induced from a single-cell E. amylovora, while no change in the control bacterial cell (Bacillus subtilis) was observed, or negligible change, if any. Furthermore, the bacterial sensor demonstrated a potential for the continuous detection of E. amylovora through simple surface regeneration, enabling its reusability. Taken together, our system has the potential to be applied in fields where early symptoms are not observed and where single-cell or ultra-sensitive detection is required, such as plant bacterial pathogen detection, foodborne pathogen monitoring and analysis, and pathogenic microbial diagnosis. [ABSTRACT FROM AUTHOR]

In this study, we developed a highly sensitive and specific bimolecular fluorescence complementation (BiFC)-based influenza A virus (IAV)-sensing system by combining a galactose/glucose-binding protein (GGBP) with an N-terminal large domain (YN1-172) and a C-terminal small domain (YC173-239) made up of enhanced yellow fluorescence protein (eYFP). The GGBP-based BiFC reporter exhibits the fluorescence reconstitution as a result of conformational changes in GGBP when lactose, which was derived from 6′-silalyllactose and used as a substrate for neuraminidase (NA), binds to GGBP in the presence of IAV. The system showed a linear dynamic range extending from 1 × 100 to 1 × 107 TCID50/mL, and it had a detection limit of 1.1 × 100 TCID50/mL for IAV (H1N1), demonstrating ultra-high sensitivity. Our system exhibited fluorescence intensity enhancements in the presence of IAV, while it displayed weak fluorescence signals when exposed to NA-deficient viruses, such as RSV A, RSV B, adenovirus and rhinovirus, thereby indicating selective responses for IAV detection. Overall, our system provides a simple, highly sensitive and specific IAV detection platform based on BiFC that is capable of detecting ligand-induced protein conformational changes, obviating the need for virus culture or RNA extraction processes. [ABSTRACT FROM AUTHOR]