Search results for: lateral flow immunoassay on-site detection

Authors: Jaeyoung Kim, Heeju Lee, Huijin Jung, Heesoo Pyo, Seungki Kim, Joonseok Lee

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Avian influenza viruses (AIV) are the primary cause of highly contagious respiratory diseases caused by type A influenza viruses of the Orthomyxoviridae family. AIV are categorized on the basis of types of surface glycoproteins such as hemagglutinin and neuraminidase. Certain H5 and H7 subtypes of AIV have evolved to the high pathogenic avian influenza (HPAI) virus, which has caused considerable economic loss to the poultry industry and led to severe public health crisis. Several commercial kits have been developed for on-site detection of AIV. However, the sensitivity of these methods is too low to detect low virus concentrations in clinical samples and opaque stool samples. Here, we introduced a background-free near-infrared (NIR)-to-NIR upconversion nanoparticle-based lateral flow immunoassay (NNLFA) platform to yield a sensor that detects AIV within 20 minutes. Ca²⁺ ion in the shell was used to enhance the NIR-to-NIR upconversion photoluminescence (PL) emission as a heterogeneous dopant without inducing significant changes in the morphology and size of the UCNPs. In a mixture of opaque stool samples and gold nanoparticles (GNPs), which are components of commercial AIV LFA, the background signal of the stool samples mask the absorption peak of GNPs. However, UCNPs dispersed in the stool samples still show strong emission centered at 800 nm when excited at 980 nm, which enables the NNLFA platform to detect 10-times lower viral load than a commercial GNP-based AIV LFA. The detection limit of NNLFA for low pathogenic avian influenza (LPAI) H5N2 and HPAI H5N6 viruses was 10² EID₅₀/mL and 10³.⁵ EID₅₀/mL, respectively. Moreover, when opaque brown-colored samples were used as the target analytes, strong NIR emission signal from the test line in NNLFA confirmed the presence of AIV, whereas commercial AIV LFA detected AIV with difficulty. Therefore, we propose that this rapid and background-free NNLFA platform has the potential of detecting AIV in the field, which could effectively prevent the spread of these viruses at an early stage.

Keywords: avian influenza viruses, lateral flow immunoassay on-site detection, upconversion nanoparticles

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Authors: Joel Ballesteros, Harriet Jane Caleja, Florian Del Mundo, Cherrie Pascual

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Diffraction-based sensing was utilized in the quantification of human ferritin in blood serum to provide an alternative to label-based immunoassays currently used in clinical diagnostics and researches. The diffraction intensity was measured by the diffractive optics technology or dotLab™ system. Two methods were evaluated in this study: direct immunoassay and direct sandwich immunoassay. In the direct immunoassay, human ferritin was captured by human ferritin antibodies immobilized on an avidin-coated sensor while the direct sandwich immunoassay had an additional step for the binding of a detector human ferritin antibody on the analyte complex. Both methods were repeatable with coefficient of variation values below 15%. The direct sandwich immunoassay had a linear response from 10 to 500 ng/mL which is wider than the 100-500 ng/mL of the direct immunoassay. The direct sandwich immunoassay also has a higher calibration sensitivity with value 0.002 Diffractive Intensity (ng mL-1)-1) compared to the 0.004 Diffractive Intensity (ng mL-1)-1 of the direct immunoassay. The limit of detection and limit of quantification values of the direct immunoassay were found to be 29 ng/mL and 98 ng/mL, respectively, while the direct sandwich immunoassay has a limit of detection (LOD) of 2.5 ng/mL and a limit of quantification (LOQ) of 8.2 ng/mL. In terms of accuracy, the direct immunoassay had a percent recovery of 88.8-93.0% in PBS while the direct sandwich immunoassay had 94.1 to 97.2%. Based on the results, the direct sandwich immunoassay is a better diffraction-based immunoassay in terms of accuracy, LOD, LOQ, linear range, and sensitivity. The direct sandwich immunoassay was utilized in the determination of human ferritin in blood serum and the results are validated by Chemiluminescent Magnetic Immunoassay (CMIA). The calculated Pearson correlation coefficient was 0.995 and the p-values of the paired-sample t-test were less than 0.5 which show that the results of the direct sandwich immunoassay was comparable to that of CMIA and could be utilized as an alternative analytical method.

Keywords: biosensor, diffraction, ferritin, immunoassay

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Authors: Wanwisa Deenin, Abdulhadee Yakoh, Chahya Kreangkaiwal, Orawon Chailapakul, Kanitha Patarakul, Sudkate Chaiyo

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LipL32 is an outer membrane protein present only on pathogenic Leptospira species, which are the causative agent of leptospirosis. Leptospirosis symptoms are often misdiagnosed with other febrile illnesses as the clinical manifestations are non-specific. Therefore, an accurate diagnostic tool for leptospirosis is indeed critical for proper and prompt treatment. Typical diagnosis via serological assays is generally performed to assess the antibodies produced against Leptospira. However, their delayed antibody response and complicated procedure are undoubtedly limited the practical utilization especially in primary care setting. Here, we demonstrate for the first time an early-stage detection of LipL32 by an integrated lateral-flow immunoassay with electrochemical readout (eLFIA). A ferrocene trace tag was monitored via differential pulse voltammetry operated on a smartphone-based device, thus allowing for on-field testing. Superior performance in terms of the lowest detectable limit of detection (LOD) of 8.53 pg/mL and broad linear dynamic range (5 orders of magnitude) among other sensors available thus far was established. Additionally, the developed test strip provided a straightforward yet sensitive approach for diagnosis of leptospirosis using the collected human sera from patients, in which the results were comparable to the real-time polymerase chain reaction technique.

Keywords: leptospirosis, electrochemical detection, lateral flow immunosensor, point-of-care testing, early-stage detection

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Authors: Xinyi Zhao, Furong Tian

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Mycotoxins are secondary metabolic products of fungi. These are poisonous, carcinogens and mutagens in nature and pose a serious health threat to both humans and animals, causing severe illnesses and even deaths. The rapid, simple and cheap detection methods of mycotoxins are of immense importance and in great demand in the food and beverage industry as well as in agriculture and environmental monitoring. Lateral flow immunochromatographic strips (ICSTs) have been widely used in food safety, environment monitoring. Forty-six papers were identified and reviewed on Google Scholar and Scopus for their limit of detection and nanomaterial on Lateral flow immunochromatographic strips on different types of mycotoxins. The papers were dated 2001-2021. Twenty five papers were compared to identify the lowest limit of detection of among different mycotoxins (Aflatoxin B1: 10, Zearalenone:5, Fumonisin B1: 5, Trichothecene-A: 5). Most of these highly sensitive strips are competitive. Sandwich structure are usually used in large scale detection. In conclusion, the mycotoxin receives that most researches is aflatoxin B1 and its limit of detection is the lowest. Gold-nanopaticle based immunochromatographic test strips has the lowest limit of detection. Five papers involve smartphone detection and they all detect aflatoxin B1 with gold nanoparticles. In these papers, quantitative concentration results can be obtained when the user uploads the photograph of test lines using the smartphone application.

Keywords: aflatoxin B1, limit of detection, gold nanoparticle, lateral flow immunochromatographic strips, mycotoxins

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Authors: Pattarachaya Preechakasedkit, Kota Osada, Koji Suzuki, Daniel Citterio, Orawon Chailapakul

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A paper-based lateral flow device for screening thyroid stimulating hormone (TSH) is reported. A sandwich immunoassay was performed using two mouse monoclonal TSH antibodies (anti-hTSH 5403 and 5404) as immobilized and labeled antibodies for capturing TSH samples. Test (anti-hTSH 5403) and control (goat anti-Mouse IgG) lines were fabricated on nitrocellulose membrane (NCM) using ballpoint pen printed with a speed of 3 cm/s and thickness setting of 1. The novel gold nanoparticles europium complex (AuNPs@Eu) was used as fluorescence label compared to conventional AuNPs label. The results obtained with this device can be visually assessed by the naked eyes and under UV hand lamps, and quantitative analysis can be performed using the ImageJ program. The limit of detection (LOD) under UV hand lamps (0.1 µIU/mL) provided 50-fold greater sensitivity than AuNPs (5 µIU/mL), which is suitable for both hypothyroidism and hyperthyroidism screening within 30 min. A linear relationship between the red intensity and the logarithmic concentrations of TSH was observed with a good correlation (R²=0.992). Furthermore, the device can be effectively applied for screening TSH in the spiked human serum with recovery range of 96.80-104.45% and RSD of 2.18-3.63%. Therefore, the developed device is an alternative method for TSH screening which provides a lot of advantages including low cost, short time analysis, ease of use, disposability, portability, and on-site measurement.

Keywords: thyroid stimulating hormone, paper-based lateral flow, hypothyroidism, hyperthyroidism

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Authors: Chang, Ya-Ju, Hsieh, Pei-Hsin, Wu, Jui-Chuang, Chen-Yang, Yui Whei

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A mesoporous silica material was prepared to apply to the lateral-flow immunochromatography for detecting a model biosample. The probe antibody is immobilized on the silica surface as the test line to capture its affinity antigen, which laterally flows through the chromatography strips. The antigen is labeled with nano-gold particles, such that the detection can be visually read out from the test line without instrument aids. The result reveals that the mesoporous material provides a vast area for immobilizing the detection probes. Biosening surfaces corresponding with a positive proportion of detection signals is obtained with the biosample loading.

Keywords: mesoporous silica, immunochromatography, lateral-flow strips, biosensors, nano-gold particles

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Authors: Mark Osborn

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Clustered, regularly interspaced short palindromic repeat (CRISPR/Cas) proteins can be designed to bind specified DNA and RNA sequences and hold great promise for the accurate detection of nucleic acids for diagnostics. Commercially available reagents were integrated into a CRISPR/Cas9-based lateral flow assay that can detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) sequences with single-base specificity. This approach requires minimal equipment and represents a simplified platform for field-based deployment. A rapid, multiplex fluorescence CRISPR/Cas9 nuclease cleavage assay capable of detecting and differentiating SARS-CoV-2, influenza A and B, and respiratory syncytial virus in a single reaction was also developed. These findings provide proof of principle for CRISPR/Cas9 point-of-care diagnosis that can detect specific SARS-CoV-2 strain(s). Further, Cas9 cleavage allows for a scalable fluorescent platform for identifying respiratory viral pathogens with overlapping symptomology. Collectively, this approach is a facile platform for diagnostics with broad application to user-defined sequence interrogation and detection.

Keywords: CRISPR/Cas9, lateral flow assay, SARS-Co-V2, single-nucleotide resolution

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Authors: M. Salvador, J. C. Martinez-Garcia, A. Moyano, M. C. Blanco-Lopez, M. Rivas

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Biosensors play a crucial role in the detection of molecules nowadays due to their advantages of user-friendliness, high selectivity, the analysis in real time and in-situ applications. Among them, Lateral Flow Immunoassays (LFIAs) are presented among technologies for point-of-care bioassays with outstanding characteristics such as affordability, portability and low-cost. They have been widely used for the detection of a vast range of biomarkers, which do not only include proteins but also nucleic acids and even whole cells. Although the LFIA has traditionally been a positive/negative test, tremendous efforts are being done to add to the method the quantifying capability based on the combination of suitable labels and a proper sensor. One of the most successful approaches involves the use of magnetic sensors for detection of magnetic labels. Bringing together the required characteristics mentioned before, our research group has developed a biosensor to detect biomolecules. Superparamagnetic nanoparticles (SPNPs) together with LFIAs play the fundamental roles. SPMNPs are detected by their interaction with a high-frequency current flowing on a printed micro track. By means of the instant and proportional variation of the impedance of this track provoked by the presence of the SPNPs, quantitative and rapid measurement of the number of particles can be obtained. This way of detection requires no external magnetic field application, which reduces the device complexity. On the other hand, the major limitations of LFIAs are that they are only qualitative or semiquantitative when traditional gold or latex nanoparticles are used as color labels. Moreover, the necessity of always-constant ambient conditions to get reproducible results, the exclusive detection of the nanoparticles on the surface of the membrane, and the short durability of the signal are drawbacks that can be advantageously overcome with the design of magnetically labeled LFIAs. The approach followed was to coat the SPIONs with a specific monoclonal antibody which targets the protein under consideration by chemical bonds. Then, a sandwich-type immunoassay was prepared by printing onto the nitrocellulose membrane strip a second antibody against a different epitope of the protein (test line) and an IgG antibody (control line). When the sample flows along the strip, the SPION-labeled proteins are immobilized at the test line, which provides magnetic signal as described before. Preliminary results using this practical combination for the detection and quantification of the Prostatic-Specific Antigen (PSA) shows the validity and consistency of the technique in the clinical range, where a PSA level of 4.0 ng/mL is the established upper normal limit. Moreover, a LOD of 0.25 ng/mL was calculated with a confident level of 3 according to the IUPAC Gold Book definition. Its versatility has also been proved with the detection of other biomolecules such as troponin I (cardiac injury biomarker) or histamine.

Keywords: biosensor, lateral flow immunoassays, point-of-care devices, superparamagnetic nanoparticles

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Authors: Anton Stadler, Thorsten Ike

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In this paper, we present a video based smoke detection algorithm based on TVL1 optical flow estimation. The main part of the algorithm is an accumulating system for motion angles and upward motion speed of the flow field. We optimized the usage of TVL1 flow estimation for the detection of smoke with very low smoke density. Therefore, we use adapted flow parameters and estimate the flow field on difference images. We show in theory and in evaluation that this improves the performance of smoke detection significantly. We evaluate the smoke algorithm using videos with different smoke densities and different backgrounds. We show that smoke detection is very reliable in varying scenarios. Further we verify that our algorithm is very robust towards crowded scenes disturbance videos.

Keywords: low density, optical flow, upward smoke motion, video based smoke detection

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Authors: Zdenek Silar, Martin Dobrovolny

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This article deals with the obstacle detection on a railway crossing (clearance detection). Detection is based on the optical flow estimation and classification of the flow vectors by K-means clustering algorithm. For classification of passing vehicles is used optical flow direction determination. The optical flow estimation is based on a modified Lucas-Kanade method.

Keywords: background estimation, direction of optical flow, K-means clustering, objects detection, railway crossing monitoring, velocity vectors

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Authors: Cristina Hotoleanu

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Introduction: The recent Covid-19 pandemic accelerated the development of e-health, including telemedicine, smartphone applications, and medical wearable devices. Providing remote teleconsultations supposes challenges which may require further face-to-face medical interactions. The aim of this study was to assess the correlation between the types of teleconsultations and the need of onsite medical services (investigations and medical visits) for the diagnosis and treatment. Methods: a retrospective study including all the teleconsultations using the platform offered by a telehealth provider in Romania (Telios Care SA) between May 1, 2021- April 30, 2022, was performed. Binary data were analysed using the chi-square test with a significance level of p < 0.05. Results: out of 7163 consultations, 3961 were phone calls, 1981 were online messages, and 1221 were video calls. Onsite medical services were indicated in 3327 (46.44%) cases; the onsite investigations or the onsite visits were recommended for 2908 patients as follows: 2326 in case of phone calls, 582 in case of online messages, none in case of video calls. Both onsite investigations and visits were indicated for 419 patients. The need for onsite additional medical services was significantly higher in the case of phone calls than in the other 2 types of teleconsultations (Chi square= 1207.06, p= 0.00001). The indication for onsite services was done mainly after teleconsultations covering medical specialties (87.34%), significantly higher than the other specialties (Chi square=914.59, p=0.00001). Teleconsultations in surgical specialties and other fields (pharmacy, dentistry, psychology, wellbeing- nutrition, fitness) resulted in 12.13%, respective less than 1%, indication for onsite investigations or visits, explained by using of video calls in most of the cases. Conclusion: a further onsite medical service was necessary in less than a half of the teleconsultations. This indication was done mainly after phone calls and teleconsultations in medical specialties. Video calls were used mostly in psychology, nutrition, and fitness teleconsultations and did not require a further onsite medical service. Other studies are necessary to assess better the types of teleconsultations and the specialties bringing the biggest benefit for the patients.

Keywords: onsite medical services, phone calls, teleconsultations, telemedicine

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Authors: Spandan Sahu, Amiya Kumar Pati, Kishanjit Kumar Khatua

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Rivers are the backbone of human civilization as well as one of the most important components of nature. In this paper, a method for predicting lateral depth-averaged velocity distribution in a two-flow braided compound channel is proposed. Experiments were conducted to study the boundary shear stress in the tip of the two flow path. The cross-section of the channel is divided into several panels to study the flow phenomenon on both the main channel and the flood plain. It can be inferred from the study that the flow coefficients get affected by boundary shear stress. In this study, the analytical solution of Shiono and knight (SKM) for lateral distributions of depth-averaged velocity and bed shear stress has been taken into account. The SKM is based on hydraulic parameters, which signify the bed friction factor (f), lateral eddy viscosity, and depth-averaged flow. While applying the SKM to different panels, the equations are solved considering the boundary conditions between panels. The boundary shear stress data, which are obtained from experimentation, are compared with CES software, which is based on quasi-one-dimensional Reynold's Averaged Navier-Stokes (RANS) approach.

Keywords: boundary shear stress, lateral depth-averaged velocity, two-flow braided compound channel, velocity distribution

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Authors: Muhammad Solikhudin Nafi, Wahyu Afif Mufida, Mita Erna Wati, Fitri Setyani Rokim, M. Al-Rizqi Dharma Fauzi

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High dose activity without any pre-exercise will impact Delayed Onset Muscle Soreness (DOMS). DOMS known as delayed pain post-exercise and induce skeletal injury which will decrease athletes’ performances. From now on, post-exercise muscle damage can be detected by measuring skeletal troponin I (sTnI) concentration in serum using ELISA but this method needs more time and cost. To prevent decreased athletes performances, screening need to be done rapidly. We want to introduce our new prototype to detect DOMS acutely. Rapid detection tests are based on immunological reaction between skeletal troponin I antibodies and sTnI in human serum or whole blood. Chemical methods that are used in the manufacture of diagnostic test is lateral flow immunoassay. The material used is rat monoclonal antibody sTnI, colloidal gold, anti-mouse IgG, nitrocellulose membrane, conjugate pad, sample pad, wick and backing card. The procedure are made conjugate (colloidal gold and mAb sTnI) and insert into the conjugate pad, gives spray sTnI mAb and anti-mouse IgG into nitrocellulose membrane, and assemble RDT. RDT had been evaluated by measuring the sensitivity of positive human serum (n = 30) and negative human serum (n = 30). Overall sensitivity value was 93% and specificity value was 90%. ADCOR as the first rapid detection test qualitatively showed antigen-antibody reaction and showed good overall performances for screening of muscle damage. Furthermore, these finding still need more improvements to get best results.

Keywords: DOMS, sTnI, rapid detection test, ELISA

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Authors: Josselyn Mata Calidonio, Arianna I. Maddox, Kimberly Hamad-Schifferli

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Rapid diagnostics are critical infectious disease tools that are designed to detect a known biomarker using antibodies specific to that biomarker. However, a way to detect unknown viruses has not yet been achieved in a paper test format. We describe here a route to make an adaptable paper immunoassay that can detect an unknown biomarker, demonstrating it on SARS-CoV-2 variants. The immunoassay repurposes cross-reactive antibodies raised against the alpha variant. Gold nanoparticles of two different colors conjugated to two different antibodies create a colorimetric signal, and machine learning of the resulting colorimetric pattern is used to train the assay to discriminate between variants of alpha and Omicron BA.5. By using principal component analysis, the colorimetric test patterns can pick up and discriminate an unknown that it has not encountered before, Omicron BA.1. The test has an accuracy of 100% and a potential calculated discriminatory power of 900. We show that it can be used adaptively and that it can be used to pick up emerging variants without the need to raise new antibodies.

Keywords: adaptive immunoassay, detecting unknown viruses, gold nanoparticles, paper immunoassay, repurposing antibodies

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Authors: Yang Yang, Dan Liu

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Anomaly detection models for network flow based on machine learning have poor detection performance under extremely unbalanced training data conditions and also have slow detection speed and large resource consumption when deploying on network edge devices. Embedding multi-teacher knowledge distillation (mKD) in anomaly detection can transfer knowledge from multiple teacher models to a single model. Inspired by this, we proposed a state-of-the-art model, mKDNAD, to improve detection performance. mKDNAD mine and integrate the knowledge of one-dimensional sequence and two-dimensional image implicit in network flow to improve the detection accuracy of small sample classes. The multi-teacher knowledge distillation method guides the train of the student model, thus speeding up the model's detection speed and reducing the number of model parameters. Experiments in the CICIDS2017 dataset verify the improvements of our method in the detection speed and the detection accuracy in dealing with the small sample classes.

Keywords: network flow anomaly detection (NAD), multi-teacher knowledge distillation, machine learning, deep learning

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Authors: Vinci Mojamdar, Govind S. Gupta

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Moving beds in the presence of gas flow are widely used in metallurgical and chemical industries like blast furnaces, catalyst reforming, drying, etc. Pressure drop studies in co- and counter – current conditions have been done by a few researchers. However, to the best of authours knowledge, proper pressure drop study with lateral gas injection lacks especially in the presence of cavity and nozzle protrusion inside the packed bed. The latter study is more useful for metallurgical industries for the processes such as blast furnaces, shaft reduction and, COREX. In this experimental work, a two dimensional cold model with slot type nozzle for lateral gas injection along with the plastic beads as packing material and dry air as gas have been used. The variation of pressure drop is recorded at various horizontal and vertical directions in the presence of cavity and nozzle protrusion. The study has been performed in both moving and stationary beds. Also, the experiments have been carried out in both increasing as well as decreasing gas flow conditions. Experiments have been performed at various gas flow rates and packed bed heights. Some interesting results have been reported such as there is no pressure variation in the moving bed for both the increasing and decreasing gas flow condition that is different from the stationary bed. Pressure hysteresis loop has been observed in a stationary bed.

Keywords: lateral gas injection, moving bed, pressure drop, pressure hysteresis, stationary bed

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Authors: Zhen Cao, Yu Zhu, Junxue Fu

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Immunoassay, i.e., antigen-antibody reaction, is crucial for disease diagnostics. To achieve the adequate signal of the antigen protein detection, a large amount of sample and long incubation time is needed. However, the amount of protein is usually small at the early stage, which makes it difficult to detect. Unlike cells and DNAs, no valid chemical method exists for protein amplification. Thus, an alternative way to improve the signal is through particle manipulation techniques to concentrate proteins, among which dielectrophoresis (DEP) is an effective one. DEP is a technique that concentrates particles to the designated region through a force created by the gradient in a non-uniform electric field. Since DEP force is proportional to the cube of particle size and square of electric field gradient, it is relatively easy to capture larger particles such as cells. For smaller ones like proteins, a super high gradient is then required. In this work, three-dimensional Ag/SiO2 nanorods arrays, fabricated by an easy physical vapor deposition technique called as oblique angle deposition, have been integrated with a DEP device and created the field gradient as high as of 2.6×10²⁴ V²/m³. The nanorods based DEP device is able to enrich bovine serum albumin (BSA) protein by 1800-fold and the rate has reached 180-fold/s when only applying 5 V electric potential. Based on the above nanorods integrated DEP platform, an immunoassay of mouse immunoglobulin G (IgG) proteins has been performed. Briefly, specific antibodies are immobilized onto nanorods, then IgG proteins are concentrated and captured, and finally, the signal from fluorescence-labelled antibodies are detected. The limit of detection (LoD) is measured as 275.3 fg/mL (~1.8 fM), which is a 20,000-fold enhancement compared with identical assays performed on blank glass plates. Further, prostate-specific antigen (PSA), which is a cancer biomarker for diagnosis of prostate cancer after radical prostatectomy, is also quantified with a LoD as low as 2.6 pg/mL. The time to signal saturation has been significantly reduced to one minute. In summary, together with an easy nanorod fabrication and integration method, this nanorods based DEP platform has demonstrated highly sensitive immunoassay performance and thus poses great potentials in applications for early point-of-care diagnostics.

Keywords: dielectrophoresis, immunoassay, oblique angle deposition, protein concentration

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Authors: Sonu Gandhi, Neena Capalash, Prince Sharma, C. Raman Suri

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A sensitive chemiluminescence immunoassay (CIA) for heroin and its major metabolites is reported. The method is based on the competitive reaction of horseradish peroxidase (HRP)-labeled anti-MAM antibody and free drug in spiked urine samples. A hapten-protein conjugate was synthesized by using acidic derivative of monoacetyl morphine (MAM) coupled to carrier protein BSA and was used as an immunogen for the generation of anti-MAM (monoacetyl morphine) antibody. A high titer of antibody (1:64,0000) was obtained and the relative affinity constant (Kaff) of antibody was 3.1×107 l/mol. Under the optimal conditions, linear range and reactivity for heroin, mono acetyl morphine (MAM), morphine and codeine were 0.08, 0.09, 0.095 and 0.092 ng/mL respectively. The developed chemiluminescence inhibition assay could detect heroin and its metabolites in standard and urine samples up to 0.01 ng/ml.

Keywords: heroin, metabolites, chemiluminescence immunoassay, horse radish peroxidase

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Authors: Najwa Mimouni, Omar Rahli, Rachid Bennacer, Salah Chikh

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In the present work 2D and 3D numerical simulations of double diffusion natural convection in an elongated enclosure filled with a binary fluid saturating a porous medium are carried out. In the formulation of the problem, the Boussinesq approximation is considered and cross Neumann boundary conditions are specified for heat and mass walls conditions. The numerical method is based on the control volume approach with the third order QUICK scheme. Full approximation storage (FAS) with full multigrid (FMG) method is used to solve the problem. For the explored large range of the controlling parameters, we clearly evidenced that the increase in the depth of the cavity i.e. the lateral aspect ratio has an important effect on the flow patterns. The 2D perfect parallel flows obtained for a small lateral aspect ratio are drastically destabilized by increasing the cavity lateral dimension. This yields a 3D fluid motion with a much more complicated flow pattern and the classically studied 2D parallel flows are impossible.

Keywords: bifurcation, natural convection, heat and mass transfer, parallel flow, porous media

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Authors: Dan W. Rhodes, Juliane Hey, Magali Karagueuzian, Florianne Martinez, Yael Sandowski, Vanessa Roulet, Mahmoud Badawi, Mohammed-Amine Chakir, Valérie Simon, Jérémie Gautier, Françoise Le Boulaire, Catherine Coignard, Claire Vincent, Sandrine Greaume, Isabelle Voisin

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Background: Beckman Coulter, Inc. (BEC) has recently developed a fully automated second-generation anti-HCV test on a new immunoassay platform. The objective of this multicenter study conducted in Europe was to evaluate the performance of the ACCESS anti-HCV assay on the recently CE-marked DxI 9000 ACCESS Immunoassay Analyzer as an aid in the diagnosis of HCV (Hepatitis C Virus) infection and as a screening test for blood and plasma donors. Methods: The clinical specificity of the ACCESS anti-HCV assay was determined using HCV antibody-negative samples from blood donors and hospitalized patients. Sample antibody status was determined by a CE-marked anti-HCV assay (Abbott ARCHITECTTM anti-HCV assay or Abbott PRISM HCV assay) with an additional confirmation method (Immunoblot testing with INNO-LIATM HCV Score - Fujirebio), if necessary, according to pre-determined testing algorithms. The clinical sensitivity was determined using known HCV antibody-positive samples, identified positive by Immunoblot testing with INNO-LIATM HCV Score - Fujirebio. HCV RNA PCR or genotyping was available on all Immunoblot positive samples for further characterization. The false initial reactive rate was determined on fresh samples from blood donors and hospitalized patients. Thirty (30) commercially available seroconversion panels were tested to assess the sensitivity for early detection of HCV infection. The study was conducted from November 2019 to March 2022. Three (3) external sites and one (1) internal site participated. Results: Clinical specificity (95% CI) was 99.7% (99.6 – 99.8%) on 5852 blood donors and 99.0% (98.4 – 99.4%) on 1527 hospitalized patient samples. There were 15 discrepant samples (positive on ACCESS anti-HCV assay and negative on both ARCHITECT and Immunoblot) observed with hospitalized patient samples, and of note, additional HCV RNA PCR results showed five (5) samples had positive HCV RNA PCR results despite the absence of HCV antibody detection by ARCHITECT and Immunoblot, suggesting a better sensitivity of the ACCESS anti-HCV assay with these five samples compared to the ARCHITECT and Immunoblot anti-HCV assays. Clinical sensitivity (95% CI) on 510 well-characterized, known HCV antibody-positive samples was 100.0% (99.3 – 100.0%), including 353 samples with known HCV genotypes (1 to 6). The overall false initial reactive rate (95% CI) on 6630 patient samples was 0.02% (0.00 – 0.09%). Results obtained on 30 seroconversion panels demonstrated that the ACCESS anti-HCV assay had equivalent sensitivity performances, with an average bleed difference since the first reactive bleed below one (1), compared to the ARCHITECTTM anti-HCV assay. Conclusion: The newly developed ACCESS anti-HCV assay from BEC for use on the DxI 9000 ACCESS Immunoassay Analyzer demonstrated high clinical sensitivity and specificity, equivalent to currently marketed anti-HCV assays, as well as a low false initial reactive rate.

Keywords: DxI 9000 ACCESS Immunoassay Analyzer, HCV, HCV antibody, Hepatitis C virus, immunoassay

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Authors: Milad Alizadeh Galdiani, Jabbar Ali Zakeri

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Lateral movement of CWR ballasted track occurs in sharp curves because of the lack of adequate lateral resistance. Several strategies have been proposed and used for increase the lateral resistance of ballasted tracks, but still there are some problems in tracks with small radius curves. In this paper, a new method has been presented for increase the lateral resistance. This method is using the lateral supports as numerical and field studies. In this paper, the field and laboratory tests have been conducted by using the single tie pressure test (STPT) and track panel loading test (LTPT). Then, their results were compared with the numerical results. The results of numerical and field tests showed that the lateral stiffness of ballasted tracks significantly increased when there were lateral supports in ballasted tracks. Also, the track structure had a bilinear behavior.

Keywords: ballasted railway, Lateral resistance, railway buckling, field and numerical studies

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Authors: Vanessa Roulet, Marc Turini, Juliane Hey, Stéphanie Bord-Romeu, Emilie Bonzom, Mahmoud Badawi, Mohammed-Amine Chakir, Valérie Simon, Vanessa Viotti, Jérémie Gautier, Françoise Le Boulaire, Catherine Coignard, Claire Vincent, Sandrine Greaume, Isabelle Voisin

Abstract:

Background: Beckman Coulter, Inc. has recently developed fully automated assays for the detection of HBsAg on a new immunoassay platform. The objective of this European multicenter study was to evaluate the performance of the ACCESS HBsAg and ACCESS HBsAg Confirmatory assays† on the recently CE-marked DxI 9000 ACCESS Immunoassay Analyzer. Methods: The clinical specificity of the ACCESS HBsAg and HBsAg Confirmatory assays was determined using HBsAg-negative samples from blood donors and hospitalized patients. The clinical sensitivity was determined using presumed HBsAg-positive samples. Sample HBsAg status was determined using a CE-marked HBsAg assay (Abbott ARCHITECT HBsAg Qualitative II, Roche Elecsys HBsAg II, or Abbott PRISM HBsAg assay) and a CE-marked HBsAg confirmatory assay (Abbott ARCHITECT HBsAg Qualitative II Confirmatory or Abbott PRISM HBsAg Confirmatory assay) according to manufacturer package inserts and pre-determined testing algorithms. False initial reactive rate was determined on fresh hospitalized patient samples. The sensitivity for the early detection of HBV infection was assessed internally on thirty (30) seroconversion panels. Results: Clinical specificity was 99.95% (95% CI, 99.86 – 99.99%) on 6047 blood donors and 99.71% (95%CI, 99.15 – 99.94%) on 1023 hospitalized patient samples. A total of six (6) samples were found false positive with the ACCESS HBsAg assay. None were confirmed for the presence of HBsAg with the ACCESS HBsAg Confirmatory assay. Clinical sensitivity on 455 HBsAg-positive samples was 100.00% (95% CI, 99.19 – 100.00%) for the ACCESS HBsAg assay alone and for the ACCESS HBsAg Confirmatory assay. The false initial reactive rate on 821 fresh hospitalized patient samples was 0.24% (95% CI, 0.03 – 0.87%). Results obtained on 30 seroconversion panels demonstrated that the ACCESS HBsAg assay had equivalent sensitivity performances compared to the Abbott ARCHITECT HBsAg Qualitative II assay with an average bleed difference since first reactive bleed of 0.13. All bleeds found reactive in ACCESS HBsAg assay were confirmed in ACCESS HBsAg Confirmatory assay. Conclusion: The newly developed ACCESS HBsAg and ACCESS HBsAg Confirmatory assays from Beckman Coulter have demonstrated high clinical sensitivity and specificity, equivalent to currently marketed HBsAg assays, as well as a low false initial reactive rate. †Pending achievement of CE compliance; not yet available for in vitro diagnostic use. 2023-11317 Beckman Coulter and the Beckman Coulter product and service marks mentioned herein are trademarks or registered trademarks of Beckman Coulter, Inc. in the United States and other countries. All other trademarks are the property of their respective owners.

Keywords: dxi 9000 access immunoassay analyzer, hbsag, hbv, hepatitis b surface antigen, hepatitis b virus, immunoassay

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Authors: Ka Ho Yau, Jan Frederick Engels, Kwok Kei Lai, Reinhard Renneberg

Abstract:

Lateral flow test is a prevalent technology in various sectors such as food, pharmacology and biomedical sciences. Colloidal gold (CG) is widely used as the signalling molecule because of the ease of synthesis, bimolecular conjugation and its red colour due to intrinsic SPRE. However, the production of colloidal gold is costly and requires vigorous conditions. The stability of colloidal gold are easily affected by environmental factors such as pH, high salt content etc. Silica nanoparticles are well known for its ease of production and stability over a wide range of solvents. Using reverse micro-emulsion (w/o), silica nanoparticles with different sizes can be produced precisely by controlling the amount of water. By incorporating different water-soluble dyes, a rainbow colour of the silica nanoparticles could be produced. Conjugation with biomolecules such as antibodies can be achieved after surface modification of the silica nanoparticles with organosilane. The optimum amount of the antibodies to be labelled was determined by Bradford Assay. In this work, we have demonstrated the ability of the dye-doped silica nanoparticles as a signalling molecule in lateral flow test, which showed a semi-quantitative measurement of the analyte. The image was further analysed for the LOD=10 ng of the analyte. The working range and the linear range of the test were from 0 to 2.15μg/mL and from 0 to 1.07 μg/mL (R2=0.988) respectively. The performance of the tests was comparable to those using colloidal gold with the advantages of lower cost, enhanced stability and having a wide spectrum of colours. The positives lines can be imaged by naked eye or by using a mobile phone camera for a better quantification. Further research has been carried out in multicolour detection of different biomarkers simultaneously. The preliminary results were promising as there was little cross-reactivity being observed for an optimized system. This approach provides a platform for multicolour detection for a set of biomarkers that enhances the accuracy of diseases diagnostics.

Keywords: colorimetric detection, immunosensor, paper-based biosensor, silica

Procedia PDF Downloads 348

Authors: Ying Li, Rui Hu, Shizhen Chen, Xin Zhou, Yunhuang Yang

Abstract:

Prostate cancer is one of the leading causes of cancer-related death among men. Prostate-specific antigen (PSA), a specific product of prostatic epithelial cells, is an important indicator of prostate cancer. Though PSA is not a specific serum biomarker for the screening of prostate cancer, it is recognized as an indicator for prostate cancer recurrence and response to therapy for patient’s post-prostatectomy. Since radical prostatectomy eliminates the source of PSA production, serum PSA levels fall below 50 pg/mL, and may be below the detection limit of clinical immunoassays (current clinical immunoassay lower limit of detection is around 10 pg/mL). Many clinical studies have shown that intervention at low PSA levels was able to improve patient outcomes significantly. Therefore, ultra-sensitive and precise assays that can accurately quantify extremely low levels of PSA (below 1-10 pg/mL) will facilitate the assessment of patients for the possibility of early adjuvant or salvage treatment. Currently, the commercially available ultra-sensitive ELISA kit (not used clinically) can only reach a detection limit of 3-10 pg/mL. Other platforms developed by different research groups could achieve a detection limit as low as 0.33 pg/mL, but they relied on sophisticated instruments to get the final readout. Herein we report a microfluidic platform for point-of-care (POC) detection of PSA with a detection limit of 0.5 pg/mL and without the assistance of any equipment. This platform is based on a previously reported volumetric-bar-chart chip (V-Chip), which applies platinum nanoparticles (PtNPs) as the ELISA probe to convert the biomarker concentration to the volume of oxygen gas that further pushes the red ink to form a visualized bar-chart. The length of each bar is used to quantify the biomarker concentration of each sample. We devised a long reading channel V-Chip (LV-Chip) in this work to achieve a wide detection window. In addition, LV-Chip employed a unique enzyme-free ELISA probe that enriched PtNPs significantly and owned 500-fold enhanced catalytic ability over that of previous V-Chip, resulting in a significantly improved detection limit. LV-Chip is able to complete a PSA assay for five samples in 20 min. The device was applied to detect PSA in 50 patient serum samples, and the on-chip results demonstrated good correlation with conventional immunoassay. In addition, the PSA levels in finger-prick whole blood samples from healthy volunteers were successfully measured on the device. This completely stand-alone LV-Chip platform enables convenient POC testing for patient follow-up in the physician’s office and is also useful in resource-constrained settings.

Keywords: point-of-care detection, microfluidics, PSA, ultra-sensitive

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Authors: Anas M. Fares

Abstract:

Reinforced concrete shear walls are the most frequently used forms of lateral resisting structural elements. These walls may take many forms due to their functions and locations in the building. In Palestine, the most lateral resisting forces construction forms is the cantilever shear walls system. It is thus of prime importance to study the rigidity of these walls. The virtual work theorem is used to derive the total lateral deflection of cantilever shear walls due to flexural and shear deformation. The case of neglecting the shear deformation in the walls is also studied, and it is found that the wall height to length aspect ratio (H/B) plays a major role in calculating the lateral deflection and the rigidity of such walls. When the H/B is more than or equal to 3.7, the shear deformation may be neglected from the calculation of the lateral deflection. Moreover, the walls with the same material properties, same lateral load value, and same aspect ratio, shall have the same of both the lateral deflection and the rigidity. Finally, an equation to calculate the total rigidity and total deflection of such walls is derived by using the virtual work theorem for a cantilever beam.

Keywords: cantilever shear walls, flexural deformation, lateral deflection, lateral loads, reinforced concrete shear walls, rigidity, shear deformation, virtual work theorem

Procedia PDF Downloads 190

Authors: Jungbhin You, Taekyun Kim, Jongho Park, Sungnam Hong, Sun-Kyu Park

Abstract:

Reinforced concrete has been mainly used in construction field because of excellent durability. However, it may lead to reduction of durability and safety due to corrosion of reinforcement steels according to damage of concrete surface. Recently, research of textile is ongoing to complement weakness of reinforced concrete. In previous research, only experiment of longitudinal length were performed. Therefore, in order to investigate the adhesion performance according to the lattice shape and the embedded length, the pull-out test was performed on the roving with parameter of the number of lateral reinforcement, the lateral reinforcement length and the lateral reinforcement spacing. As a result, the number of lateral reinforcement and the lateral reinforcement length did not significantly affect the load variation depending on the adhesion performance, and only the load analysis results according to the reinforcement spacing are affected.

Keywords: adhesion performance, lateral reinforcement, pull-out test, textile

Procedia PDF Downloads 335

Authors: Xinghang Lv, Tao Peng, Jia Chen, Junping Liu, Xinrong Hu, Ruhan He, Minghua Jiang, Wenli Cao

Abstract:

As one of the most dangerous vulnerabilities, effective detection of buffer overflow vulnerabilities is extremely necessary. Traditional detection methods are not accurate enough and consume more resources to meet complex and enormous code environment at present. In order to resolve the above problems, we propose the method for Buffer overflow detection based on Abstract syntax tree, Control flow graph, and Data dependency graph (BodeACD) in C/C++ programs with source code. Firstly, BodeACD constructs the function samples of buffer overflow that are available on Github, then represents them as code representation sequences, which fuse control flow, data dependency, and syntax structure of source code to reduce information loss during code representation. Finally, BodeACD learns vulnerability patterns for vulnerability detection through deep learning. The results of the experiments show that BodeACD has increased the precision and recall by 6.3% and 8.5% respectively compared with the latest methods, which can effectively improve vulnerability detection and reduce False-positive rate and False-negative rate.

Keywords: vulnerability detection, abstract syntax tree, control flow graph, data dependency graph, code representation, deep learning

Procedia PDF Downloads 142

Authors: Hyungsuk Han, Soohong Jeon, Chungwon Lee, YongHoon Kim

Abstract:

When a rubber mount and flexible coupling are installed on the main engine, high torsional vibration can occur. The root cause of this high torsional vibration can be attributed to the lateral-torsional coupled vibration of the shaft system. Therefore, the lateral-torsional coupled vibration is investigated numerically after approximating the shaft system to a three-degrees-of-freedom Jeffcott rotor. To verify that the high torsional vibration is caused by the lateral-torsional coupled vibration, a test unit that can simulate this lateral-torsional coupled vibration occurring in the propulsion shaft is developed. Performing a vibration test with the test unit, it can be experimentally verified that the high torsional vibration occurring in the propulsion shaft of the particular ship was caused by the lateral-torsional coupled vibration.

Keywords: Jeffcott rotor, lateral-torsional coupled vibration, propulsion shaft, stability

Procedia PDF Downloads 204

Authors: Yeongyu Choi, Ju H. Park, S. M. Lee, Ho-Youl Jung

Abstract:

In this paper, an improved method for estimating fundamental matrix is proposed. The method is applied effectively to monocular camera based moving object detection. The method consists of corner points detection, moving object’s motion estimation and fundamental matrix calculation. The corner points are obtained by using Harris corner detector, motions of moving objects is calculated from pyramidal Lucas-Kanade optical flow algorithm. Through epipolar geometry analysis using RANSAC, the fundamental matrix is calculated. In this method, we have improved the performances of moving object detection by using two threshold values that determine inlier or outlier. Through the simulations, we compare the performances with varying the two threshold values.

Keywords: corner detection, optical flow, epipolar geometry, RANSAC

Procedia PDF Downloads 377

Authors: Xavier A. Conlan

Abstract:

Speed of analysis is a significant limitation to current high-performance liquid chromatography/mass spectrometry (HPLC/MS) and ultra-high-pressure liquid chromatography (UHPLC)/MS systems both of which are used in many forensic investigations. The flow rate limitations of MS detection require a compromise in the chromatographic flow rate, which in turn reduces throughput, and when using modern columns, a reduction in separation efficiency. Commonly, this restriction is combated through the post-column splitting of flow prior to entry into the mass spectrometer. However, this results in a loss of sensitivity and a loss in efficiency due to the post-extra column dead volume. A new chromatographic column format known as 'parallel segmented flow' involves the splitting of eluent flow within the column outlet end fitting, and in this study we present its application in order to interrogate the provenience of methamphetamine samples with mass spectrometry detection. Using parallel segmented flow, column flow rates as high as 3 mL/min were employed in the analysis of amino acids without post-column splitting to the mass spectrometer. Furthermore, when parallel segmented flow chromatography columns were employed, the sensitivity was more than twice that of conventional systems with post-column splitting when the same volume of mobile phase was passed through the detector. These finding suggest that this type of column technology will particularly enhance the capabilities of modern LC/MS enabling both high-throughput and sensitive mass spectral detection.

Keywords: chromatography, mass spectrometry methamphetamine, parallel segmented outlet flow column, forensic sciences

Procedia PDF Downloads 460