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The fast development of the Internet of things (IoT) promises to deliver convenience to human life. However, a huge amount of the data is constantly generated, transmitted, processed, and stored, posing significant security challenges. The currently available security protocols and encryption techniques are mostly based on software algorithms and pseudorandom number generators that are vulnerable to attacks. A true random number generator (TRNG) based on devices using stochastically physical phenomena has been proposed for auditory data encryption and trusted communication. In the current study, a Bi

Recently, the deep learning (DL) dimension of artificial intelligence has received much attention from biochemical researchers and thus has gradually become the key approach adopted in the area of biosensing applications. Studies have shown that the use of DL techniques for sensing can not only shorten the time of data analysis but also significantly increase the accuracy of data analysis and prediction, resulting in the performance improvement of biosensing systems in comparison to conventional methods. However, obtaining reliable equilibrium and rate constants of biomolecular interactions during the detection process remains difficult and time-consuming to date. In this study, we propose a transformed model based on the deep transfer learning and sequence-to-sequence autoencoder that can successfully transfer the SPR sensorgram to the protein-binding constants, that is, the association rate constant (

Background: preterm and critically ill neonates often experience clinically suspected sepsis during their prolonged hospitalization in the neonatal intensive care unit (NICU), which can be the initial sign of final adverse outcomes. Therefore, we aimed to utilize machine learning approaches to predict neonatal in-hospital mortality through data-driven learning. Methods: a total of 1095 neonates who experienced clinically suspected sepsis in a tertiary-level NICU in Taiwan between August 2017 and July 2020 were enrolled. Clinically suspected sepsis was defined based on clinical features and laboratory criteria and the administration of empiric antibiotics by clinicians. The variables used for analysis included patient demographics, clinical features, laboratory data, and medications. The machine learning methods used included deep neural network (DNN), k-nearest neighbors, support vector machine, random forest, and extreme gradient boost. The performance of these models was evaluated using the area under the receiver operating characteristic curve (AUC). Results: the final in-hospital mortality of this cohort was 8.2% (90 neonates died). A total of 765 (69.8%) and 330 (30.2%) patients were randomly assigned to the training and test sets, respectively. Regarding the efficacy of the single model that most accurately predicted the outcome, DNN exhibited the greatest AUC (0.923, 95% confidence interval [CI] 0.953–0.893) and the best accuracy (95.64%, 95% CI 96.76–94.52%), Cohen’s kappa coefficient value (0.74, 95% CI 0.79–0.69) and Matthews correlation coefficient value (0.75, 95% CI 0.80–0.70). The top three most influential variables in the DNN importance matrix plot were the requirement of ventilator support at the onset of suspected sepsis, the feeding conditions, and intravascular volume expansion. The model performance was indistinguishable between the training and test sets. Conclusions: the DNN model was successfully established to predict in-hospital mortality in neonates with clinically suspected sepsis, and the machine learning algorithm is applicable for clinicians to gain insights and have better communication with families in advance.

Cytomegalovirus (CMV) is the most frequent cause of congenital infection worldwide; congenital CMV may lead to significant mortality, morbidity, or long-term sequelae, such as sensorineural hearing loss. The current study presents a newly designed surface plasmon resonance (SPR) biosensor for CMV-specific microRNAs that does not involve extra care for receptor immobilization or treatment to prevent fouling on bare gold surfaces. The modification-free approach, which utilizes a poly-adenine [poly(A)]-Au interaction, exhibited a high affinity that was comparable to that of the gold-sulfur (Au-S) interaction. In addition, magnetic nanoparticles (MNPs) were used to separate the analyte from complex sample matrixes that significantly reduced nonspecific adsorption. Moreover, the MNPs also played an important role in SPR signal amplification due to the binding-induced change in the refractive index. Our SPR biosensing platform was used successfully for the multi-detection of the microRNAs, UL22A-5p, and UL112-3p, which were associated with CMV. Our SPR biosensor offered the detection limits of 108 fM and 24 fM for UL22A-5p and UL112-3p, respectively, with an R2 of 0.9661 and 0.9985, respectively. The precision of this biosensor has an acceptable CV (coefficient of variation) value of

Background Areas of hypoxia are often found in triple-negative breast cancer (TNBC), it is thus more difficult to treat than other types of breast cancer, and may require combination therapies. A new strategy that combined bioreductive therapy with photodynamic therapy (PDT) was developed herein to improve the efficacy of cancer treatment. Our design utilized the characteristics of protoporphyrin IX (PpIX) molecules that reacted and consumed O2 at the tumor site, which led to the production of cytotoxic reactive oxygen species (ROS). The low microenvironmental oxygen levels enabled activation of a bioreductive prodrug, tirapazamine (TPZ), to become a toxic radical. The TPZ radical not only eradicated hypoxic tumor cells, but it also promoted therapeutic efficacy of PDT. Results To achieve the co-delivery of PpIX and TPZ for advanced breast cancer therapy, thin-shell hollow mesoporous Ia3d silica nanoparticles, designated as MMT-2, was employed herein. This nanocarrier designed to target the human breast cancer cell MDA-MB-231 was functionalized with PpIX and DNA aptamer (LXL-1), and loaded with TPZ, resulting in the formation of TPZ@LXL-1-PpIX-MMT-2 nanoVector. A series of studies confirmed that our nanoVectors (TPZ@LXL-1-PpIX-MMT-2) facilitated in vitro and in vivo targeting, and significantly reduced tumor volume in a xenograft mouse model. Histological analysis also revealed that this nanoVector killed tumor cells in hypoxic regions efficiently. Conclusions Taken together, the synergism and efficacy of this new therapeutic design was confirmed. Therefore, we concluded that this new therapeutic strategy, which exploited a complementary combination of PpIX and TPZ, functioned well in both normoxia and hypoxia, and is a promising medical procedure for effective treatment of TNBC.

Mycobacterium tuberculosis complex (MTBC) is the well-known causative agents of tuberculosis (TB). Transmission of bacteria via airborne routes makes infection control difficult. Hence, rapid and accurate identification of TB from clinical sputum samples is essential for minimizing the spread of the disease. However, traditional bacterial identification methods for TB are time consuming and labor intensive. The aim of this study is to propose a straightforward and rapid screening platform for MTBC using a portable organic light-emitting diode- (OLED-) based surface plasmon resonance (SPR) biosensor integrated with a nested PCR technique. Experimentally, the limit of detection (LOD) for MTBC PCR products measurement was estimated to be around 63 pg/mL. In addition, the biosensor can successfully differentiate MTBC from other nontuberculous mycobacterium strains. In 600 clinical sputum specimens, the sensitivity and specificity were 96.6% (86/89) and 98.4% (503/511), respectively. These results indicate that the portable OLED-based SPR biosensor may be taken into account as a suitable and convenient tool employed in the remote area and developing country for diagnosing TB diseases.

AIM To investigate the feasibility of reducing the scan time of paediatric technetium 99m (99mTc) dimercaptosuccinic acid (DMSA) single-photon-emission computed tomographic (SPECT) using a deep learning (DL) method. MATERIAL AND METHODS A total of 112 paediatric 99mTc-DMSA renal SPECT scans were analysed retrospectively. Of the 112 examinations, 88 (84 for training and four for validation) were used to train a DL-based model that could generate full-acquisition-time reconstructed SPECT images from half-time acquisition. The remaining 24 examinations were used to evaluate the performance of the trained model. RESULTS DL-based SPECT images obtained from half-time acquisition have image quality similar to the standard clinical SPECT images obtained from full-acquisition-time acquisition. Moreover, the accuracy, sensitivity and specificity of the DL-based SPECT images for detection of affected kidneys were 91.7%, 83.3%, and 100%, respectively. CONCLUSION These preliminary results suggest that DL has the potential to reduce the scan time of paediatric 99mTc-DMSA SPECT imaging while maintaining diagnostic accuracy.

Tyramine (4-hydroxyphenethylamine), which is a monoamine metabolized by monoamine oxidase (MAO), exists widely in plants, animals, fermented foods, and salted foods. The incidence of hypertension, or "cheese effect", which is associated with a large dietary intake of tyramine while taking MAO inhibitors has been reported; therefore, the measurement of tyramine is an urgent concern. Herein, an efficient approach that integrates a molecular imprinting polymer for solid phase extraction (MISPE) technique with a sensitive electrochemical sensing platform (SPCE/PEDOT: PSS/AuNP/1-m-4-MP) for the quantification of tyramine is presented. Enhanced electrode conductivity was achieved sequentially by constructing a conductive polymer (PEDOT: PSS) on a screen-printed carbon electrode (SPCE), followed by electrodeposition with gold nanoparticles (AuNPs) and, finally, by modification with positively charged 1-methyl-4-mercaptopyridine (1-m-4-MP) using an Au-S bond. Tyramine was isolated selectively and pre-concentrated by the MISPE technique; electroanalysis that used differential pulse voltammetry (DPV) in NaOH (0.1M, pH 13) was conducted successively. Experimental parameters (such as modes of electrode modification, ratio of PEDOT: PSS, pH of electrolyte, time required for AuNP deposition, and 1-m-4-MP concentrations) that were associated with optimal detection conditions were evaluated also. We obtained a linear concentration range (5-100nM, R2=0.9939) with LOD and sensitivity at 2.31nM, and 3.11μAnM-1cm-2, respectively. The applicability of our technique was demonstrated by analyzing tyramine in spiked serum and milk. The feature of our newly developed analytical methods that coupled sample pre-treatment (sample clean-up and pre-concentration) with sensitive detection makes it a promising tool for quantifying of tyramine.

Lung cancer is the primary cause of cancer-associated mortality worldwide, which makes the identification of reliable lung cancer biomarkers a pressing need for early diagnosis and prognosis. RGS11, which is a regulator of G-protein signaling and also a lung cancer biomarker, plays an important role in cancer-related metastasis. However, trace levels of RGS11 (in the range of pg/mL) in serum samples make it difficult to quantify using currently available enzyme-linked immunosorbent assay (ELISA) kits and, therefore, this hinders progress in the discovery of new approaches for treating lung cancer. The aim of this study is to develop a rapid, sensitive, and reliable platform for the detection of RGS11 lung cancer biomarker based on a suspension immunoassay coupled with an isothermal exponential amplification strategy. Our study was initiated by the functionalization of magnetic beads with anti-RGS11 antibodies (Ab-MB) by EDC (1-ethyl-3-(3-(dimethylamino)propyl)-carbodiimide)/NHS ( N-hydroxysulfosuccinimide) activation. Ab-MB served as a sensing probe for the competitive immunorecognitions between known concentrations of His-tag RGS11 and unknown concentrations of target RGS11 in serum. The reporter anti-His antibodies, which were modified with primers that induced an isothermal exponential amplification reaction, were subsequently introduced to the reaction mixture that resulted in the formation of immunosandwich complexes. The exponentially amplified DNA duplex that was intercalated with SYBR Green was designated as a signal reporter for the assessment of RGS11 in an inversely proportional relationship. The sensing platform was excellent for the determination of RGS11 with an exceptional detection limit of 148 fg/mL and a linear dynamic range of 0.1-10 pg/mL using a minimal sample volume (20 μL) and with a reaction time of 1.5 h. In addition, we challenged the sensing platform with RGS11-spiked samples (in 2× diluted serum), and an acceptable recovery rate (90%) was observed. Finally, 24 clinical samples acquired from patients with advanced lung cancer (C), inflammation (I), and heart failure (H) were analyzed by this newly developed sensing platform and a commercial ELISA kit for validation. This sensing platform has potential in biomedical applications for clinically diagnosing liquid biopsy samples for patients with lung cancer. Moreover, the universal design of our proposed system is easily adapted to detect any other protein if a His-tag recombinant protein is available.

In this study, we designed a four-layered (Al/Au/Ag/Au) sensor chip for surface plasmon resonance (SPR) biosensor and demonstrated that the structure shows a better performance than conventional SPR chip in our system.

A white organic light-emitting diode (OLED) with enhanced sensitivity has been demonstrated as a novel light source in a portable surface plasmon resonance (SPR) optical biosensor. A disposable broad-spectral OLED was employed on the leg side of isosceles trapezoid prism for a fixed angle of incident light. Bimetallic Ag/Au composition layers were used as sensing layers and their structures evaluated such that the wavelength resonance occurred at the peak of the light spectrum. The SPR signal detection applied differential intensities at two reference wavelengths. We show that the integration of a white spectral OLED on an SPR sensor improved the sensor's sensitivity by ∼19.29% compared to an SPR sensor system using a green OLED for a bimetallic Ag/Au sensing layer. The limit of detection (LOD) of 2.4 × 10−6 refractive index units (RIU) has been established in the range of refractive index samples (Δn) around 3.6 × 10−3 RIU. This optical sensor demonstrated the capability of rapid, real-time monitoring operation. The activation of self-assembled monolayer (SAM), antibody immobilization, and biomolecular interaction detection of immunoglobulin G (IgG) protein are recorded successfully with a detection limit around 40.3 pg/mL.

In 2013 a new reassortant avian influenza A H7N9 virus emerged in China, causing human infection with high mortality. An accurate and timely diagnosis is crucial for controlling the outbreaks of the disease. We therefore propose a simple strategy for rapidly and sensitively detecting the H7N9 virus using an intensity-modulated surface plasmon resonance (IM-SPR) biosensor integrated with a new generated monoclonal antibody. The novel antibody exhibits significant specificity to recognize H7N9 virus compared with other clinical human influenza isolates (p0.01). Experimentally, the detection limit of the proposed approach for H7N9 virus detection is estimated to be 144 copies/mL, which is a 20-fold increase in sensitivity compared with homemade target-captured ELISA using the identical antibody. For the measurement of mimic clinical specimens containing the H7N9 virus mixed with nasal mucosa from flu-like syndrome patients, the detection limit is calculated to be 402 copies/mL, which is better than conventional influenza detection assays; quantitative reverse transcription polymerase chain reaction (qRT-PCR) and rapid influenza diagnostic test (RIDT). Most importantly, the assay time took less than 10 min. Combined, the results of this study indicate that the proposed simple strategy demonstrates high sensitivity and time-saving in H7N9 virus detection. By incorporating a high specific recognizer, the proposed technique has the potential to be used in applications and development of other emerging or re-emerging microbe detection platforms.

Human metapneumovirus (hMPV) is a common respiratory tract infection in children. However, conventional immunofluorescence assays (IFAs) for detecting hMPV in respiratory samples have limited reliability with a sensitivity and false-negative predictive value of 58.1% and approximately 17.8%, respectively. In this study, hMPV was measured in 91 clinical respiratory samples (55 sputum and 36 nasopharyngeal aspirate samples), which were obtained from children under three years of age, utilizing our previously developed high-throughput metal-enhanced fluorescence (MEF)-based biosensor (HT-MEFB). The sensitivity of HT-MEFB for hMPV detection in the 91 samples was improved by up to 77.4% compared with that obtained with IFAs, and the specificity of HT-MEFB for hMPV detection was 91.7%. In addition, the specificity and accuracy obtained after the selection of 55 sputum samples as the analyzed specimen reached 92.3% and 90.9%, respectively. Thus, in terms of accuracy, high throughput, and sensitivity, HT-MEFB exhibits considerable potential for hMPV detection in clinical settings.

This study aims to improve the signal sensitivity of a portable surface plasmon resonance (SPR) sensor system. An organic light emitting diode (OLED) was integrated with a brightness enhancement film (BEF) and a giant birefringent optical (GBO) film as a light source to construct the SPR based on an OLED–BEF–GBO film. A method for calculating the SPR signal by summing the differences in optical intensity at two different wavelengths was used to minimize the effect of the noise error that particularly occurs in a portable SPR device. The experimental results indicated that the limit of detection (LOD) of the SPR using an Au sensing layer has an effective refractive index change of 7.8 × 10−6 RIU. Furthermore, for the SPR using Au/Ag sensing metal layers, an LOD of 3.2 × 10−6 RIU was achieved due to the improved sensitivity and resolution value compared with the use of an Au sensing layer. The use of the SPR device in a bio-affinity assay to test the interaction between goat anti-mouse immunoglobulin G (IgG) and mouse IgG protein achieved a LOD of approximately 40.6 pg/mL.

White-light scanning interferometry (WLSI) is often used to study the surface profiles and properties of thin films because the strength of the technique lies in its ability to provide fast and high resolution measurements. An innovative attempt is made in this paper to apply WLSI as a time-domain spectroscopic system for localized surface plasmon resonance (LSPR) sensing. A WLSI-based spectrometer is constructed with a breadboard of WLSI in combination with a spectral centroid algorithm for noise reduction and performance improvement. Experimentally, the WLSI-based spectrometer exhibits a limit of detection (LOD) of 1.2 × 10−3 refractive index units (RIU), which is better than that obtained with a conventional UV-Vis spectrometer, by resolving the LSPR peak shift. Finally, the bio-applicability of the proposed spectrometer was investigated using the rs242557 tau gene, an Alzheimer’s and Parkinson’s disease biomarker. The LOD was calculated as 15 pM. These results demonstrate that the proposed WLSI-based spectrometer could become a sensitive time-domain spectroscopic biosensing platform.

A novel “degree of coherence paired surface plasma wave biosensor” (DOC-PSPWB) is proposed wherein the principle of detection involves the degree of coherence of highly correlated surface plasma waves when these are excited on a metal/dielectric interface within the DOC-PSPWB. Using the sensor, the concentration of total PSA (t-PSA) in a phosphate-buffered saline solution was measured, and a detection limit of 0.015pg/mL was determined experimentally. Finally, the dynamic range of the DOC-PSPWB when used on samples with ultralow molecular concentrations is discussed.

Ischemic shock and acute kidney injury (AKI) are the primary causes of high mortality in the intensive care unit (ICU) patients. In a rat model, ischemia is able to disrupt the actin cytoskeleton of proximal tubule cells. This effect is associated with the expression and excretion of actin depolymerizing factor (ADF)/cofilin. However, the human evidence of ADF/cofilin excreted in ICU patients with ischemic shock and/or AKI remains to be addressed. Here we developed a 96-well high-throughput, localized surface plasmon-coupled fluorescence biosensor (HT-LSPCFB) combining a sandwich immunoassay to measure the urine cofilin-1 in 57 ICU patients and 8 healthy controls. A linear relationship between 1 and 10 5 pg/mL of human cofilin-1 recombinant protein was determined ( R 2 = 0.9845) in this study. The highest normalized cofilin-1 levels obtained in the ICU patients and healthy adults were 1.985 and 0.726, respectively. The mean normalized cofilin-1 level of total ICU patients (0.7403) was also significantly higher than that of healthy adults (0.4759) ( p = 0.0052). An examination of the receiver operating characteristic (ROC) curve and area under curve (AUC) showed that cofilin-1 is acceptable for assessing the ICU patients (AUC = 0.775) and shock (AUC = 0.713), but not for AKI (AUC = 0.681). Therefore, the HT-LSPCFB is suitable for detecting the urine cofilin-1 which potentially becomes a prognostic factor for ICU patients with shock.

In this study, we applied the developed paired surface plasma waves biosensor (PSPWB) in a dual-channel biosensor for rapid and sensitive detection of swine-origin influenza A (H1N1) virus (S-OIV). In conjunction with the amplitude ratio of the signal and the reference channel, the stability of the PSPWB system is significantly improved experimentally. The theoretical limit of detection (LOD) of the dual-channel PSPWB for S-OIV is 30 PFU/mL (PFU, plaque-forming unit), which was calculated from the fitting curve of the surface plasmon resonance signal with a S-OIV clinical isolate concentration in phosphate-buffered saline (PBS) over a range of 18-1.8 × 10(6) PFU/mL. The LOD is 2 orders of magnitude more sensitive than the commercial rapid influenza diagnostic test at worst and an order of magnitude less sensitive than real-time quantitative polymerase chain reaction (PCR) whose LOD for S-OIV in PBS was determined to be 3.5 PFU/mL in this experiment. Furthermore, under in vivo conditions, this experiment demonstrates that the assay successfully measured S-OIV at a concentration of 1.8 × 10(2) PFU/mL in mimic solution, which contained PBS-diluted normal human nasal mucosa. Most importantly, the assay time took less than 20 min. From the results, the dual-channel PSPWB potentially offers great opportunity in developing an alternative PCR-free diagnostic method for rapid, sensitive, and accurate detection of viral pathogens with epidemiological relevance in clinical samples by using an appropriate pathogen-specific antibody.

A multiplexed biosensor based on WLSI is capable to measure various gold nanoparticles accompanied by the LSPR extinction spectrum shift simultaneously. In this paper, the multiplex detection can be realized by the WLSI system.

A novel fiber-optic biosensor based on a localized surface plasmon coupled fluorescence (LSPCF) system is proposed and developed. This biosensor consists of a biomolecular complex in a sandwich format ofantibody/antigen/Cy5-antibody-gold nanoparticle (GNP). It is immobilized on the surface of an optical fiber where aCy5-antibody-GNPcomplex forms the fluorescence probe and is produced by mixing Cy5-labeled antibody and protein A conjugated gold nanoparticles (Au-PA). The LSPCF is excited by localized surface plasmon on the GNP surface where the evanescent field is applied near the core surface of the optical fiber. At the same time, the fluorescence signal is detected by a photomultiplier tube located beside the unclad optical fiber with high collection efficiency. Experimentally, this novel LSPCF biosensor is able to detect mouse immunoglobulin G (IgG) at a minimum concentration of 1 pg/mL (7 fM) during the biomolecular interaction of the IgG with anti-mouse IgG. The analysis is expanded by a discussion of the amplification of the LSPCF intensity by GNP coupling, and overall, this LSPCF biosensor is confirmed experimentally as a biosensor with very high sensitivity.

Evanescent-wave excited fluorescence technology has been demonstrated to enhance sensitivity and reduce matrix effects, making it suitable for biosensor development. In this study, we developed a liposome-based, total internal reflection fluorescence, fiber-optic biosensor (TIRF-FOB) for protein detection, which integrates a liposomal amplifier and sandwich immunoassay format with TIRF-FOB. In addition, the antibody-tagged and fluorophore-entrapped liposomes for heterogeneous detection of target molecules were designed and synthesized. This biosensor successfully detected the target protein (model analyzed here is IgG) with a limit of detection (LOD) of 2.0 attomoles for the target protein (equivalent to 2.0 pg/mL of protein presented in 150 μL of sample solution). The features of this ultra-sensitive liposomal TIRF-FOB are (i) fluorescence is excited via evanescent waves and amplified via liposomes; (ii) the use of two polyclonal antibodies in the sandwich assay format increases the specificity and lowers the cost of our assay. Based on the exceptional detection sensitivity and cost-effectiveness, we believe that the proposed biosensor has great potential as a practical, clinical diagnostic tool in the near future.

This paper describes an eco-friendly, one-pot strategy for the synthesis of water-soluble, high-quantum-yield gold nanoclusters (AuNCs) stabilized with 11-mercaptoundecanoic acid (MUA) on their surfaces. The as-prepared ultrasmall MUA-AuNCs (1.9 nm) exhibited a quantum yield (QY) of 13%, higher than those of most previously described thiol-protected AuNCs. We applied these MUA-AuNCs as a versatile probe to develop a fluorescence "turn-off" assay for sensing Hg(2+) ions as well as a fluorescence "turn-on" assay for sensing biothiols. The former assay operated through aggregation-induced fluorescence quenching upon interaction of the MUA-AuNCs with Hg(2+) ions in a buffer containing 2,6-pyridinedicarboxylic acid (PDCA); this probe provided high sensitivity and remarkable selectivity over other selected metal ions with a limit of detection (LOD) for Hg(2+) ions of 450 pM and linearity from 2 to 50 nM. In the latter assay for biothiols [i.e., cysteine (Cys), homocysteine (Hcy), glutathione (GSH)], the fluorescence of the Hg(2+)-MUA-AuNCs complexes was turned on because the affinity of Hg(2+) ions toward the SH group of the biothiols was greater than that toward the COOH groups of the MUA units on the surface of the AuNCs. This assay provided good linearity for the tested biothiols, ranging from 10 to 100 nM for Cys, from 10 to 100 nM for Hcy, and from 5 to 75 nM for GSH, with LODs of 5.4, 4.2, and 2.1 nM, respectively. In addition, these environmentally and biologically friendly AuNC probes tested satisfactorily against interference from a range of amino acids.

The actin-depolymerizing factor (ADF)/cofilin protein family has been reported to be associated with ischemia-induced renal disorders. We examine whether cofilin-1 is associated with acute kidney injury (AKI) using human urine samples. We exploited a 96-well based high-throughput biosensor that uses gold nanoparticles and a sandwich immunoassay to detect the urine cofilin-1 level of AKI patients. The mean urine cofilin-1 level of the AKI patients (n=37 from 47 cases analyzed) was twofold higher than that of healthy adults (n=21 from 29 cases analyzed). The receiver operating characteristic (ROC) curve showed that cofilin-1 was acceptable for discriminating AKI patients from healthy adults. However, an increase of the sample size is required to conclude the importance of urine cofilin-1 on AKI diagnosis, and the high-throughput ultrasensitive biosensor used in this study would greatly accelerate the measurement of urine cofilin-1 in an increased sample size.

In renal transplant patients, immunosuppressive therapy may result in the reactivation of polyomavirus BK (BKV), leading to polyomavirus-associated nephropathy (PVAN), which inevitably causes allograft failure. Since the treatment outcomes of PVAN remain unsatisfactory, early identification and continuous monitoring of BKV reactivation and reduction of immunosuppressants are essential to prevent PVAN development. The present study demonstrated that the developed dual-channel heterodyne-based surface plasmon resonance (SPR) biosensor is applicable for the rapid detection of urinary BKV. The use of a symmetrical reference channel integrated with the poly(ethylene glycol)-based low-fouling self-assembled monolayer to reduce the environmental variations and the nonspecific noise was proven to enhance the sensitivity in urinary BKV detection. Experimentally, the detection limit of the biosensor for BKV detection was estimated to be around 8500 copies/mL. In addition, urine samples from five renal transplant patients were tested to rapidly distinguish PVAN-positive and PVAN-negative renal transplant patients. By virtue of its simplicity, rapidity, and applicability, the SPR biosensor is a remarkable potential to be used for continuous clinical monitoring of BKV reactivation.

The actin depolymerizing factor (ADF)/cofilin protein family has been reported to be associated with ischemia induced renal disorders. Here we examine if cofilin-1 is associated with acute kidney injury (AKI). We exploited a 96-well based fiber-optic biosensor that uses conjugated gold nanoparticles and a sandwich immunoassay to detect the urine cofilin-1 level of AKI patients. The mean urine cofilin-1 level of the AKI patients was two-fold higher than that of healthy adults. The receiver operating characteristic (ROC) curve showed that cofilin-1 is a potential biomarker for discriminating AKI patients from healthy adults for intensive care patients.

Degree of coherence (DOC) of a paired surface plasma waves (PSPWs) in the paired surface plasma waves biosensor (PSPWB) is proposed and discussed in which a paired of surface plasma waves are excited by using a pair of highly spatial and temporal correlated P-polarized waves in a SPR device of the Kretschmann configuration. The heterodyne signal from reflected paired P-polarized laser beam is generated where the visibility of the signal is proportional to DOC of PSPWs in term of the ratio of AC and DC components of the signal. The experimental result shows that the DOC of PSPWs versus incident angle of laser beam which relates to intrinsic phonon distribution in metal film becomes much sensitive than conventional amplitude or intensity sensitive surface plasmon resonance (SPR) biosensor. Finally, the dynamic range of protein-protein interaction at ultralow concentrations is discussed.

In this study, a novel high-throughput biosensor based on metal-enhanced fluorescence technique and harmonic intensity-modulated fluorescence technique was developed and demonstrated to be highly sensitive for the detection of a pancreatic cancer marker, UL16-binding protein 2 (ULBP2), in diluted serum. Experimentally, the biosensor is able to detect ULBP2 at 16-18 pg/mL in 1% BSA-PBS and in 10-fold-diluted human serum. Compared with the limit of detection (LOD) of the conventional enzyme-linked immunosorbent assay (ELISA) method, the LOD of the proposed biosensor for ULBP2 is significantly improved by 100-fold under the same conditions. In addition, the proposed method uses two identical polyclonal antibodies for the sandwich immunoassay, simplifying the experiment in terms of the reagents needed. Consequently, this biosensor is a cost-effective tool for clinical diagnosis. We believe that the proposed high-throughput biosensor has great potential to become a clinical diagnostic tool for the detection of a pancreatic cancer marker in the near future.

Prostate-specific antigen (PSA) has been reported to be a potential biomarker of breast cancer. Serum PSA of normal women is around 1 pg/mL, which is usually undetectable by current assay methods; thus an ultrasensitive measurement of PSA expression in women's serum is necessary to distinguish normal from malignant breast diseases. To enhance the sensitivity of conventional immunoassay technology for the detection of PSA in sera, we adopted a localized surface plasmon coupled fluorescence fiber-optic biosensor, which combines a sandwich immunoassay with the localized surface plasmon technique. The concentration of total PSA (t-PSA) (from 0.1 to 1000 pg/mL) in phosphate-buffered saline solution and the normalized fluorescence signal exhibit a linear relationship where the correlation coefficient is 0.9574. In addition, the concentration of additional t-PSA in 10-fold-diluted healthly women's serum across a similar range was measured. The correlation coefficient for this measurement is 0.9142. In clinical serum samples, moreover, the experimental results of t-PSA detection show that both the mean value and median of normalized fluorescence signals in the breast cancer group (155.2 and 145.7, respectively) are higher than those in the noncancer group (46.6 and 37.1, respectively). We also examined the receiver operating characteristic curve for t-PSA, and the area under the curve (AUC) is estimated to be 0.9063, the AUC being used to measure the performance of a test to correctly identify diseased and nondiseased subjects.

Measuring the kinetic constants of protein-protein interactions at ultralow concentrations becomes critical in characterizing biospecific affinity, and exploring the feasibility of clinical diagnosis with respect to detection sensitivity, efficiency and accuracy. In this study, we propose a method that can calculate the binding constants of protein-protein interactions in sandwich assays at ultralow concentrations at the pg/mL level, using a localized surface plasmon coupled fluorescence fiber-optic biosensor (LSPCF-FOB). We discuss a two-compartment model to achieve reaction-limited kinetics under the stagnant conditions of the reaction chamber. The association rate constant, dissociation rate constant, and the equilibrium dissociation constant, that is, k(a), k(d), K(D), respectively, of the kinetics of binding between total prostate-specific antigen (t-PSA) and anti-t-PSA at concentrations from 0.1 pg/mL to 1 ng/mL, were measured either in PBS or in human serum. This is the first time that k(a), k(d), and K(D) have been measured at such a low concentration range in a complex sample such as human serum.

In this study, we describe a novel method for analyzing protein-protein binding kinetics at ultra-low concentration (1 pg/mL) using a localized surface plasmon coupled fluorescence fiber-optic biosensor (LSPCF-FOB). The association and dissociation rate constants, ka and kd, respectively, for the binding kinetics of the mouse IgG/ anti-mouse IgG interaction have been calculated to be ka = (5.9928±3.1540)x106 M-1s-1 and kd = (1.0587±0.5572)x10-3 s-1. The theoretical basis of this analytical approach is a rapid-mixing model integrated with a two-compartment model; has been experimentally verified in this study as well. The LSPCF-FOB provides a potentially alternative option for characterizing the interaction of biomolecules at ultra-low concentrations.

In order to enhance the sensitivity of conventional rapid test technique for the detection of swine-origin influenza A (H1N1) viruses (S-OIVs), we used a paired surface plasma waves biosensor (PSPWB) based on SPR in conjunction with an optical heterodyne technique. Experimentally, PSPWB showed a 125-fold improvement at least in the S-OIV detection as compared to conventional enzyme linked immunosorbent assay. Moreover, the detection limit of the PSPWB for the S-OIV detection was enhanced 250-fold in buffer at least in comparison with that of conventional rapid influenza diagnostic test.

In this study, we demonstrated that an amplitude-sensitive paired surface plasma wave biosensor (PSPWB) is capable of real-time detection of prostate-specific antigen (PSA) in diluted human serum without labeling. Experimentally, the detection limit of PSPWB was 8.4 x 10(-9) refractive index unit (RIU) and the PSPWB could measure PSA in a phosphate buffered saline solution from 10 fg/mL ( approximately 300 aM) to 100 pg/mL ( approximately 3 pM) successfully, with demonstration of a linear relationship between PSA concentrations and surface plasmon resonance (SPR) signals. Therefore, results were obtained over a wide dynamic range 5 orders of magnitude for analyte concentration. In addition, the PSPWB successfully detected PSA in diluted human serum as well. These experimental results indicate that the PSPWB is capable of detection with high sensitivity over a wide range by using SPR-based biosensors and has a capability of detecting biological analytes in clinical sample without complicated operating procedures.

The current practice for cellulitis in diagnosis and treatment is mainly based on subjective clinical judgement without validated objective guidance. For patients with non-purulent cellulitis needing intravenous antibiotic treatment in hospital, we found soft tissue sonography performed around 4 days after initiation of antibiotics might have prognostic values. The patients with soft tissue sonographic pattern of subcutaneous thickening alone had shorter duration of antibiotic treatment and higher rate of early treatment response to antibiotics than those with the pattern of cobblestone appearance. Larger-scale research may be warranted to validate the prognostic roles of sonography in cellulitis management.

Detection of severe acute respiratory syndrome (SARS) coronavirus nucleocapsid protein in human serum using localized surface plasmon coupled fluorescence (LSPCF) fiber-optic biosensor. The detection limit at 1pg/mL in human serum is successfully demonstrated.

Prostate-specific antigen (PSA) in diluted human serum was detected by using a novel amplitude-sensitive paired surface plasma waves biosensor (PSPWB). The detection limit on PSA concentration in diluted human serum at 61pg/mL (2pM) was demonstrated.

Fiber-optic biosensors have been studied intensively because they are very useful and important tools for monitoring biomolecular interactions. Here we describe a fluorescence detection fiber-optic biosensor (FD–FOB) using a sandwich assay to detect antibody–antigen interaction. In addition, the quantitative measurement of binding kinetics, including the association and dissociation rate constants for immunoglobulin G (IgG)/anti-mouse IgG, is achieved, indicating 0.38 × 10 6 M −1 s −1 for k a and 3.15 × 10 −3 s −1 for k d . These constants are calculated from the fluorescence signals detected on fiber surface only where the excited evanescent wave can be generated. Thus, a confined fluorescence-detecting region is achieved to specifically determine the binding kinetics at the vicinity of the interface between sensing materials and uncladded fiber surface. With this FD–FOB, the mathematical deduction and experimental verification of the binding kinetics in a sandwich immunoassay provide a theoretical basis for measuring rate constants and equilibrium dissociation constants. A further measurement to study the interaction between human heart-type fatty acid-binding protein and its antibody gave the calculated kinetic constants k a , k d , and K D as 8.48 × 10 5 M −1 s −1 , 1.7 × 10 −3 s −1 , and 2.0 nM, respectively. Our study is the first attempt to establish a theoretical basis for the florescence-sensitive immunoassay using a sandwich format. Moreover, we demonstrate that the FD–FOB as a high-throughput biosensor can provide an alternative to the chip-based biosensors to study real-time biomolecular interaction.

In this paper, a novel differential-phase-sensitive surface plasmon resonance biosensor (DP-SPRB) is proposed and developed, in which a two-frequency laser is integrated with a differential amplifier in order to analytically convert the phase modulation into amplitude modulation. With the use of the conventional envelope detection technique, the differential phase is precisely decoded in real time in terms of the demodulated amplitude. In order to verify high detection sensitivity of the DP-SPRB, a sucrose-water solution and glycerin-water solution at low concentrations were both tested, and the experimental results confirm that the detection sensitivity on wt % concentration of the sucrose solution is 0.00001%. Moreover, the real-time monitoring mouse IgG/antimouse IgG interaction shows the minimum concentration of mouse IgG to be at 10 fg/mL. To our knowledge, this is the highest sensitivity ever measured by a surface plasmon resonance biosensor. However, because of the limited dynamic range of DP-SPRB, it can only apply to biomolecule interactions at extremely low concentration.

In this study, we demonstrated that the fiber-optic biosensor based on localized surface plasmon coupled fluorescence (LSPCF) is capable of detecting alpha-fetoprotein (AFP) in human serum. The sensitivity of LSPCF fiber-optic biosensor is not only enhanced but also the specific selectivity is improved since the fluorophores are excited by the localized surface plasmon with high efficiency. Experimentally, this fiber-optic biosensor is able to detect AFP concentration in phosphate buffered saline (PBS) solution from 0.1ng/mL to 100ng/mL whereas the linear relationship between the AFP concentrations and the fluorescence signals is shown. Furthermore, a linear response between the fluorescence signals and the concentrations of AFP in human serum from 2.33ng/mL to 143.74ng/mL is also obtained. As a result, the detection limit of the LSPCF fiber-optic biosensor on AFP detection is comparable with the conventional enzyme-linked immunosorbent assay (ELISA). Additionally, the LSPCF fiber-optic biosensor benefits on inexpensive, disposable and simpler optical geometry that can become a high efficient immunoassay comparable with the conventional ELISA and radioimmunoassay (RIA) clinically.

Alpha-fetoprotein (AFP) detection by using a localized surface plasmon coupled fluorescence (LSPCF) fiber-optic biosensor is setup and experimentally demonstrated. It is based on gold nanoparticle (GNP) and coupled with localized surface plasmon wave on the surface of GNP. In this experiment, the fluorophores are labeled on anti-AFP which are bound to protein A conjugated GNP. Thus, LSPCF is excited with high efficiency in the near field of localized surface plasmon wave. Therefore, not only the sensitivity of LSPCF biosensor is enhanced but also the specific selectivity of AFP is improved. Experimentally, the ability of real time measurement in the range of AFP concentration from 0.1ng/ml to 100ng/ml was detected. To compare with conventional methods such as enzyme-linked immunosorbent assay (ELISA) or radioimmunoassay (RIA), the LSPCF fiber-optic biosensor performs higher or comparable detection sensitivity, respectively.

A polarized photon-pairs heterodyne polarimetry is proposed in order to measure in an ultrasensitive manner the circular birefringence of a chiral medium via optical rotation detection. A balanced detector is integrated into this polarimeter. Thus, shot-noise-limited detection by this polarimeter can be achieved. Experimentally, the detection sensitivity for the circular birefringence of a glucose-water solution up to partial differential |n(r) - n(l)| = 2 x 10(-11) at 10 mg/dL is verified. To our knowledge, this is the highest sensitivity ever measured of a chiral liquid solution based on single traveling sample cell geometry. Finally, when compared to a fiber loop ring-resonator in the frequency domain for a chiral liquid, this polarimeter shows an order of 10(4) enhancement on the sensitivity of natural optical activity measurement.

We proposed and developed a novel fiber-optic biosensor based on localized surface plasmon coupled emission (LSPCE) which consists of sandwich format of immuno-complex. It is immobilized on the surface of optical fiber where is a fluorescence probe produced by mixing Cy5 labeled antibody and protein A conjugated gold nanoparticles (Au-PA). The fluorophores are excited by localized surface plasmon (LSP) on gold nanoparticle (GNP) surface where the evanescent field is applied near the core surface of unclad optical fiber. Meanwhile, the fluorescence signal is detected by a photomultiplier tube being set beside the unclad optical fiber with high collection efficiency. In the experiment, this novel LSPCE biosensor demonstrates the minimum detectable concentration of mouse immunoglobulin G (IgG) at 1pg/ml (7fM) in the biomolecular interaction with anti-mouse IgG. From the experimental result, it verifies that LSPCE biosensor is a very high sensitive biosensor which is capable of measuring biomolecular interaction at very low concentration.