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MicroRNAs (miRNAs) have emerged as a promising class of biomarkers for early detection of various cancers, including ovarian cancer. However, quantifying miRNAs in human blood samples is challenging owing to the issues of sensitivity and specificity. In this study, hsa-miR-200a-3p of the miR-200a sub-family, which is a biomarker of ovarian cancer, was used as the analyte to demonstrate the analytical capability of an integrated biosensing platform using an extremely sensitive surface-enhanced Raman scattering (SERS) nanotag−nanoaggregate-embedded beads (NAEBs), magnetic nanoparticles (MNPs), a pair of highly specific locked nucleic acid (LNA) probes, and a semi-automated paper-based electrowetting-on-dielectric (pEWOD) device to provide labor-less and thorough sample cleanup and recovery. A sandwich approach where NAEBs are modified by one LNA-1 probe and MNPs are modified by another LNA-2 probe was applied. Then, the target analyte miRNA-200a-3p was introduced to form a sandwich nanocomplex through hybridization with the pair of LNA probes. The pEWOD device was used to achieve short cleanup time and good recovery of the nanocomplex, bringing the total analysis time to less than 30 min. The detection limit of this approach can reach 0.26 fM through SERS detection. The versatility of this method without the need for RNA extraction from clinical samples is expected to have good potential in detecting other miRNAs. [ABSTRACT FROM AUTHOR]

An effective bio-sensing platform that would meet the criteria of rapid, simple, and sensitive detection is crucial to translate bench research to clinical applications. However, simultaneously rapid and sensitive biosensing remains challenging for practical biomedical applications. In this study, for the first time, we demonstrate a cost-effective, label-free, real-time, and sensitive slab waveguide-based particle plasmon resonance (WGPPR) biosensor for practical clinical applications. A suspended glass slab waveguide structure with excellent optical confinement properties was designed and fabricated as the biosensor. Gold nanoparticles (AuNPs) were deposited on the top surface of the waveguide layer to significantly enhance the optical near field through the localized surface plasmon resonance (LSPR) effect. When light travels through the waveguide, the change in the local refractive index (RI) near the surface of the AuNPs can be transformed into changes in the intensity of transmitted light, thereby enabling sensitive and real-time detection. The RI sensing experiment shows a good sensor resolution of 1.43 × 10−4 RIU, which represents a 395% enhancement compared to that of the sensor without AuNPs. Through biochemical detection experiments, we measured IgG and determined the detection limit (LOD) at 614 ng mL−1 in ∼4 min, thereby proving the feasibility of the bio-detection sensing functionality. This study demonstrates a new type of WGPPR biosensor, which offers several unique advantages such as simple structure, high sensitivity, and rapid bio-sensing for practical bio-medical sensing applications. The new biosensor also fulfils point-of-care (POC) requirements. [ABSTRACT FROM AUTHOR]

We developed a fast (<20 min), label-free fiber optic particle plasmon resonance (FOPPR) immunosensing method to detect nervous necrosis virus (NNV), which often infects high-value economic aquatic species, such as grouper. Using spiked NNV particles in a phosphate buffer as samples, the standard calibration curve obtained was linear (R2 = 0.99) and the limit of detection (LOD) achieved was 2.75 × 104 TCID50/mL, which is superior to that obtained using enzyme-linked immunosorbent assay (ELISA). By using an enhancement method called fiber optic nanogold-linked immunosorbent assay (FONLISA), the LOD can be further improved to <1 TCID50/mL, which is comparable to that found by the conventional qPCR method. Employing the larvae homogenate samples of NNV-infected grouper, the results obtained by the FOPPR biosensor agree with those obtained by the quantitative polymerase chain reaction (qPCR) method. We also examined pond water samples from an infected container in an indoor aquaculture facility. The lowest detectable level of NNV coat protein was found to be 0.17 μg/mL, which is one order lower than the LOD reported by ELISA. Therefore, we demonstrated the potential of the FOPPR biosensor as an outbreak surveillance tool, which is able to give warning indication even when the trend of larvae death toll increment is still not clear. [ABSTRACT FROM AUTHOR]

The global pandemic of COVID-19 has created an unrivalled need for sensitive and rapid point-of-care testing (POCT) methods for the detection of infectious viruses. For the novel coronavirus SARS-CoV-2, the nucleocapsid protein (N-protein) is one of the most abundant structural proteins of the virus and it serves as a useful diagnostic marker for detection. Herein, we report a fiber optic particle plasmon resonance (FOPPR) biosensor which employed a single-stranded DNA (ssDNA) aptamer as the recognition element to detect the SARS-CoV-2 N-protein in 15 min with a limit of detection (LOD) of 2.8 nM, meeting the acceptable LOD of 106 copies/mL set by the WHO target product profile. The sensor chip is a microfluidic chip based on the balance between the gravitational potential and the capillary force to control fluid loading, thus enabling the power-free auto-flowing function. It also has a risk-free self-contained design to avoid the risk of the virus leaking into the environment. These findings demonstrate the potential for designing a low-cost and robust POCT device towards rapid antigen detection for early screening of SARS-CoV-2 and its related mutants. [ABSTRACT FROM AUTHOR]

The N-terminal pro-brain natriuretic peptide (NT-proBNP) is considered an important blood biomarker for heart failure. Herein, we report about a fiber optic nanogold-linked immunosorbent assay (FONLISA) method for the rapid, sensitive, and low-cost detection of NT-proBNP. The method is based on a sandwich immunoassay approach that uses two monoclonal NT-proBNP antibodies, a capture antibody (AbC), and a detection antibody (AbD). AbD is conjugated to a free gold nanoparticle (AuNP) to form the free AuNP@AbD conjugate, and AbC is immobilized on an unclad segment of an optical fiber. The detection of analyte (A), in this case NT-proBNP, is based on the signal change due to the formation of an AuNP@AbD–A–AbC complex on the fiber core surface, where a green light transmitted through the optical fiber will decrease in intensity due to light absorption by AuNPs via the localized surface plasmon resonance effect. This method provides a wide linear dynamic range of 0.50~5000 pg·mL−1 and a limit of detection of 0.058 pg·mL−1 for NT-proBNP. Finally, the method exhibits good correlation (r = 0.979) with the commercial central laboratory-based electrochemiluminescent immunoassay method that uses a Roche Cobas e411 instrument. Hence, our method is potentially a suitable tool for point-of-care testing. [ABSTRACT FROM AUTHOR]

This article reports the use of ac-field-induced charges at the comers of microstructures on fiber-optic sensing chips to generate electro-osmotic vortex flows in flow cell channels that can accelerate solute binding on the fiber. The sensing chip made of a cyclic olefin copolymer COC substrate contained a flow cell channel of dimensions 15 mm x 1 mm x 1 mm. A partially unclad optical fiber was placed within the channel. Relief-like strip sWuctures of 25-[micro]m thickness fabricated on the channel bottom were produced with an injection-molding process. The external electric field lines penetrating through the comers of the plastic microstructures induce charges on the comer surfaces to build up electrical double layers. When a high0-frequency ac field (~100 kHz) is used to flip the field polarities quickly, neutralization of the induced charge cannot be accomplished. The electrical double layer is therefore sustained. When absorbed charges in the double layer are driven by the external field, electro-osmotic flows are generated. The unclad portion of the fiber was coated with biotin-functionalized gold nanopartcles. The streptavidin solution was filled in the channel from the feeding tube, and the ac field (~50 V/cm) was subsequently turned on for 30 s. The ac-field-induced electro-osmotic flows can accelerate solute transport in the sensing channel to enhance the binding kinetics of streptavidin molecules with biotin probes implanted on the gold nanoparticle surface. As a result, the fiber-optic localized plasmon resonance (FO-LPR) sensing signal becomes steady as soon as the external field is turned off. In contrast, the signal cannot reach steady state until 200-300 s in a typical static sensing cell. A significant reduction in the sensing response time is demonstrated. The binding assay of streptavidin with immobilized biotin on gold nanoparticle-coated sensing fibers was validated using this mixing device. The detection limit for streptavidin of [~10.sup.-11] M is close to the reported values obtained using static cells. Similarly, the sensing response time of an orchid Odontoglossum ringspot virus (ORSV) sample was reduced from 1000 to 330 s 10.1021/ac902682k

A novel organically modified silica material has been prepared by covalent linking of the carboxylic acid group of lactate dehydrogenase to the amino group of an organoalkoxysilane precursor via a carbodiimide coupling reaction during the sol-gel process. The material was used to fabricate a leak-free biosensor. The experimental variables and characteristics of the biosensors were studied by electrochemical methods. Results showed that the coenzyme concentration, mediator concentration, and electrode rotation speed will influence the sensitivity of the biosensor. The biosensors exhibited high sensitivity (1.47 [micro]A/mM), low detection limit (1.5 [micro]M), reasonable apparent activity (1.31 [min.sup.-1]), good fabrication reproducibility, and good long-term operational stability (~1 week).

A novel class of fiber-optic evanescent-wave sensor was constructed on the basis of modification of the unclad portion of an optical fiber with serf-assembled gold colloids. The optical properties and, hence, the attenuated total reflection spectrum of serf-assembled gold colloids on the optical fiber changes with different refractive index of the environment near the colloidal gold surface. With sucrose solutions of increasing refractive index, the sensor response decreases linearly. The colloidal gold surface was also functionalized with glycine, succinic acid, or biotin to enhance the selectivity of the sensor. Results show that the sensor response decreases linearly with increasing concentration of each analyte. When the colloidal gold surface was functionalized with biotin, the detection limit of the sensor for streptavidin was 9.8 x [10.sup.-11] M. Using this approach, we demonstrate proof-of-concept of a class of refractive index sensor that is sensitive to the refractive index of the environment near the colloidal gold surface and, hence, is suitable for label-free detection of molecular or biomolecular binding at the surface of gold colloids.

We have developed a gold nanoparticle‐based surface‐assisted laser desorption/ionization mass spectrometric (SALDI‐MS) method for the analysis of amphetamine‐type stimulants (ATS), including methamphetamine (MA), 3,4‐methylenedioxy‐N‐methamphetamine (MDMA), 3,4‐methylenedioxy‐N‐ethylamphetamine (MDEA), and 4‐fluoromethamphetamine (4‐FMA), by derivatization to form quaternary ammonium species. After derivatization, the ion signal intensities of the derivatives of MA, MDMA, MDEA, and 4‐FMA significantly increase as compared to that without derivatization. Moreover, there is no analyte suppression effect in our samples after derivatization. The limits of detection (LODs) for MA, MDMA, MDEA, and 4‐FMA are 0.1, 1.0, 91, and 44 ppb, respectively, with the correlation coefficients of these regressions in the calibration curves higher than 0.99. The optimal experimental conditions, including concentration of gold nanoparticles and sample preparation method, were then applied to urine samples. After a simple sample pretreatment, including extraction and derivatization, results show that the average recoveries of the SALDI‐MS method for MA and MDMA are 100.87 ± 5.8% and 98.28 ± 2.9%, respectively. In summary, we have demonstrated a method to improve the sensitivity of the SALDI method for determination of ATS by derivatization and the method has been validated in the determination of MA and MDMA in urine samples. [ABSTRACT FROM AUTHOR]

A common strategy to improve the sensitivity of a biosensor for the detection of a low abundance analyte is to preconcentrate the analyte molecules before detection. A dual-functional gold–iron oxide core–satellite hybrid nanoparticle structure is proposed in this work to overcome the drawbacks of traditional sample pretreatment methods and the methods using non-magnetic nanomaterials for sample pretreatment. The new dual-functional hybrid nanoparticle structure can simultaneously serve as a signal reporter of a biorecognition event and a preconcentrator of a target at an extremely low concentration in a nanoplasmonic biosensor. By utilizing a fiber optic nanogold-linked sorbent assay in the fiber optic particle plasmon resonance (FOPPR) biosensor and an arbitrary DNA sequence as a target, we have demonstrated that the use of the new hybrid nanoparticle structure with magnetic preconcentration improves the limit of detection (LOD) for the DNA by 18 times as compared to the same method without magnetic preconcentration, so that the LOD for detecting the DNA can be as low as 2.6 fM. The new hybrid nanoparticle structure is easy to prepare and its use in the high-sensitivity and low-cost FOPPR biosensor provides vast opportunities in point-of-care applications. [ABSTRACT FROM AUTHOR]

Mercury ion (Hg2+) is extremely toxic even at very low concentrations and its detection mainly relies on bulky and high-cost analytical instruments. Here, we introduce a fast and ultrasensitive biosensing method developed by the integration of fiber optic particle plasmon resonance and a highly selective molecular beacon with a stem-loop DNA structure immobilized on the unclad surface of an optical fiber. In the presence of Hg2+ ions and a free assisting DNA probe, the stem-loop opens and forms thymine–Hg2+–thymine complexes. A DNA reporting probe conjugated to gold nanoparticles is used as a label to interrogate the recognition event by binding with the terminal DNA binding domain of the opened stem-loop. The method provides a wide linear range of at least 5 orders from 10–14 to 10–9 M and an extremely low detection limit of 4.37 fM (0.876 ppq). It takes less than 15 min to analyze the concentration of Hg2+ ions in aqueous samples. The method is desirable for point-of-use applications because of its low-cost instrumentation and sensor chips, easy operation, and portability. [ABSTRACT FROM AUTHOR]

The major challenge in photothermal therapy (PTT) is to develop nanocomposites that simultaneously exhibit bioimaging and PTT under a single near-infrared (NIR) irradiation with high therapeutic efficiency. Herein, we present a multifunctional nanocomposite synthesized by linking NaYF4:Yb3+,Er3+ upconversion nanoparticles (UCNPs) with gold nanorods (AuNR) to exhibit fluorescence labeling, local temperature sensing and photothermal functions simultaneously with a single NIR laser excitation. The AuNR-NaYF4:Yb3+,Er3+ nanocomposite particles displayed better photothermal properties compared with pure AuNRs or a blend of AuNRs and NaYF4:Yb3+,Er3+ UCNPs. The temperature-dependent upconversion luminescence (UCL) property was used to determine local temperature at the nanocomposite particles, which is useful for selecting appropriate irradiation dosage for PTT. The therapeutic performance of the nanocomposites in PTT for OML-1 oral cancer cells was determined. For cell labeling, we successfully labeled streptavidin-linked nanocomposite particles on the surface of OML-1 oral cancer using anti-human epidermal growth factor receptor 2 (anti-Her2) antibody. Finally, the nanocomposite particles caused exceptional destruction of cancer cells up to 70% dead cells under 976 nm laser irradiation for only one min at 0.3 W/cm2 which is below the maximal permissible exposure of human skin. [ABSTRACT FROM AUTHOR]

Identification of snake venoms is a vital step in the treatment of fatal snakebites. In this study, we use the gold–thiolate interaction between a cysteine residue and gold nanoparticles to establish a SERS method for the differentiation of the venoms of Trimeresurus stejnegeri and Bungarus multicinctus. We confirm the preference of gold nanoparticles over silver for the SERS study of snake venoms by a binding experiment that also functions to differentiate the two venom samples by colorimetry and UV-vis spectroscopy. We report the SERS spectra of Trimeresurus stejnegeri and Bungarus multicinctus venoms for the first time. The spectra display distinct SERS signatures of the snake venoms on bone-shaped gold nanoparticles made with a house recipe. These signatures correlate to selected segments of the venom proteins due to the anchoring effect of the gold-cysteine bond. The method is quick as it accomplishes in situ isolation of the structure of interest to avoid tedious purification of the samples. The location of the interactive cysteine residue makes a novel characteristic of proteins in general. [ABSTRACT FROM AUTHOR]

A novel optical immunosensor for the screening of ampicillin (Amp) residues has been developed. The biosensor is based on fiber optic particle plasmon resonance detection and uses an enhancement method called as fiber optic nanogold-linked immunosorbent assay (FONLISA) for the sensitive detection of antibiotics. A commercial antibody which had a higher affinity for ampicillin than for other β-lactam antibiotics was chosen. A surface competitive binding assay was used in which a fixed concentration of antibiotic-conjugated gold nanoparticles (AuNPs) competes with free unlabeled antibiotic molecules to measure the amount of binding with antibody molecules immobilized on an optical fiber. The synthesis of the 11-mercaptoundecanoic acid (MUA)-ampicillin conjugate facilitates the attachment of the Amp molecules to AuNPs via MUA which acts as a linker between them. This AuNP–Amp conjugate was then used for the detection of β-lactam antibiotics. The practical limit of detection obtained for Amp was 0.74 ppb (7.4 × 10−10 g/mL) which is lower than the recommended maximum residue limit (MRL) for β-lactams. The method also shows a wide linear range of 4 orders. Its applicability to the determination of ampicillin in spiked milk samples has been demonstrated with good recovery and reproducibility. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRNAs) are small noncoding single-stranded ribonucleic acid molecules. This type of endogenous oligonucleotide could be secreted into the circulation and exist stably. The detection of specific miRNAs released by cancer cells potentially provides a noninvasive means to achieve early diagnosis and prognosis of cancers. However, the typical concentration of miRNAs in blood is below the ultratrace level. This study uses a simple thermoplastic microfluidic concentration device based on an ion concentration polarization mechanism to perform enrichment and cleanup and Raman sensing beads to determine miRNA quantitatively. One sample solution containing target miRNA molecules having been hybridized with two nucleotide probes, where one probe is on a Raman tag of a nanoaggregate embedded bead (NAEB) and the other probe is on a magnetic nanoparticle (MNP), is first filled into the device. When an external field is applied across a cation exchange membrane stationed in the middle conduit of the device, the MNP-miRNA-NAEB complexed particles are enriched near the membrane edge of the cathode side. The concentrated complexed particles are further trapped using an external magnet to perform washing steps to remove excess noncomplexed NAEBs. When cleanup steps are accomplished, the remaining complexed particles are loaded into one detection capillary to acquire Raman signals from the sensing beads. Compared with that using a conventional magnetic trapping device, the cleanup time is shortened from nearly an hour to less than 10 min. Sample loss during the washing steps becomes more controllable, resulting in adequate standard curve linearity (R > 0.99) ranging from 1 to 100 pM. [ABSTRACT FROM AUTHOR]

Carboxybetaine (CB)-based zwitterionic materials have attracted considerable attention due to their dual antifouling and functionalizable properties. In this communication, the elimination effect on the antifouling and pH-responsive properties of CB materials was investigated. We synthesize β- and α-substituted methyl CB materials to investigate the occurrence of elimination in the ethylene intercharge arm in a harsh basic solution. This work provides molecular understanding of a structure–property relationship of the CB moiety for material development. [ABSTRACT FROM AUTHOR]

Gastric adenocarcinoma, a single heterogeneous disease with multiple epidemiological and histopathological characteristics, accounts for approximately 10% of cancers worldwide. It is categorized into four histological types: papillary adenocarcinoma (PAC), tubular adenocarcinoma (TAC), mucinous adenocarcinoma (MAC), and signet ring cell adenocarcinoma (SRC). Effective differentiation of the four types of adenocarcinoma will greatly improve the treatment of gastric adenocarcinoma to increase its five-year survival rate. We reported here the differentiation of the four histological types of gastric adenocarcinoma from the molecularly structural viewpoint of confocal Raman microspectroscopy. In total, 79 patients underwent laparoscopic or open radical gastrectomy during 2008–2011: 21 for signet ring cell carcinoma, 21 for tubular adenocarcinoma, 14 for papillary adenocarcinoma, 6 for mucinous carcinoma, and 17 for normal gastric mucosas obtained from patients underwent operation for other benign lesions. Clinical data were retrospectively reviewed from medical charts, and Raman data were processed and analyzed by using principal component analysis (PCA) and linear discriminant analysis (LDA). Two-dimensional plots of PCA and LDA clearly demonstrated that the four histological types of gastric adenocarcinoma could be differentiated, and confocal Raman microspectroscopy provides potentially a rapid and effective method for differentiating SRC and MAC from TAC or PAC. [ABSTRACT FROM AUTHOR]