scholarly journals Gold nanoparticle-based lateral flow kit for in vitro detection of malaria antibodies

2018 ◽  
Author(s):  
◽  
Christian Lungani Mthembu
Keyword(s):  
Test Line ◽  
Point Of Care ◽  
Three Dimensional ◽  
Lateral Flow ◽  
Igg Antibody ◽  
Diagnostic Kits ◽  
Test Kit ◽  
Malaria Antigen ◽  
And Performance

This study involves the development of three-dimensional dual lateral flow diagnostic assays. These assays were fabricated with quick response (QR) barcodes to ease the accessibility and transfer test data. The assays were designed to also improve the collection and transfer of survey from point-of-care facilities to centralized laboratories, thus, these would help to speed-up response to disease out-break. The study introduces the fabrication of two barcode based malaria diagnostic in the field of diagnostics. Two lateral flow kits were modified with two QR barcodes and three QR barcodes encoded with Google analytics codes for the detection and real-time tracking of malaria lateral flow which was designed to detect Plasmodium lactate dehydrogenase (pLDH). The fabrication of test kit was achieved by attaching two and three QR barcodes into two different test kits which were encoded with websites that were linked to Google analytics website as a tracking and performance monitor. Gold nanoparticles (AuNPs) were used as a substrate, where optical and structural properties were studied using UV/Visible spectroscopy, fluorescence spectroscopy, and transmission electron microscopy (TEM). The anti-mouse IgG antibody was used as a secondary antibody to act as control and the anti-(pLDH) was stripped on the test line. Phosphate buffer was used as a mobile phase solution. The antibody binding with pLDH antigen showed red test line indicating a positive test. Two diagnostic kits for rapid detection of pLDH were developed and validated for the detection of malaria antigen with lowest detectable recombinant concentration of 10 ng.mL-1. The diagnostic kits were incorporated with two and three optimally angled QR barcodes for identifying positive and negative. The second three QR barcode embedded test kit identified positive, negative and invalid using tracked website. These QR barcodes enabled massive results and tracking with precise location of the test through Google Analytics.

scholarly journals COMPARISON OF PERFORMANCE CHARACTERISTICS BETWEEN LATERAL FLOW, ELISA AND ELECTROCHEMILUMINESCENCE IMMUNOASSAYS FOR THE DETECTION OF SARS-COV-2 ANTIBODIES AMONG HEALTHCARE WORKERS

2021 ◽  
Author(s):  
M R Shincy ◽  
Vandana Govindan ◽  
H H Sudhakar ◽  
V T Venkatesha ◽  
K Padmapriya ◽  
...  
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
Healthcare Workers ◽  
Care Pathway ◽  
Point Of Care ◽  
Lateral Flow ◽  
Central Research ◽  
Serum Samples ◽  
Igg Antibody ◽  
Human Igg ◽  
Serological Tests

ABSTRACTBackgroundMedical professionals and researchers have been urging the need for wide and rapid testing of citizens in order to plan measures that can contain the spread of the virus. Antibody tests play an important role throughout the patient care pathway and are vital for the management and surveillance of the virus. Although RT-PCR is considered as the gold standard, serological tests based on antibodies are helpful for on-time detection. We performed one to one assessment of point-of-care lateral flow assay (POCTs), enzyme immunoassay (EIAs), electrochemiluminescence immunoassay (CLIA), to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) IgG antibody.Materials and Methods611 healthcare workers were recruited between November and December 2020 at Central Research Laboratory, KIMS. Collected serum samples were analysed according to manufacturer’s protocol. The Standard Q IgG/IgM combo assay, Anti-SARS CoV-2 Human IgG ELISA, and the Elecsys® to measure the IgG titer of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).ResultsThe kits displayed a sensitivity of 61.2%,79.5%, 91.8% and specificity of 61.7%,64.1%,80.2% for the Standard Q IgG/IgM combo assay, Anti-SARS CoV-2 Human IgG ELISA, and the Elecsys® in order.ConclusionOur results indicate high sensitivity and specificity for the Elecsys® assay compared to Anti-SARS CoV-2 Human IgG ELISA, the Standard Q IgG/IgM combo assay.

scholarly journals PCB-Based Magnetometer as a Platform for Quantification of Lateral-Flow Assays

Sensors ◽  
2019 ◽  
Vol 19 (24) ◽  
pp. 5433 ◽  
Author(s):  
Mohammad Khodadadi ◽  
Long Chang ◽  
João R. C. Trabuco ◽  
Binh V. Vu ◽  
Katerina Kourentzi ◽  
...  
Keyword(s):  
High Precision ◽  
Fe3o4 Nanoparticles ◽  
Printed Circuit Board ◽  
Test Line ◽  
Point Of Care ◽  
Low Cost ◽  
Superparamagnetic Iron Oxide ◽  
High Sensitivity ◽  
Lateral Flow ◽  
Circuit Board

This work presents a proof-of-concept demonstration of a novel inductive transducer, the femtoMag, that can be integrated with a lateral-flow assay (LFA) to provide detection and quantification of molecular biomarkers. The femtoMag transducer is manufactured using a low-cost printed circuit board (PCB) technology and can be controlled by relatively inexpensive electronics. It allows rapid high-precision quantification of the number (or amount) of superparamagnetic nanoparticle reporters along the length of an LFA test strip. It has a detection limit of 10−10 emu, which is equivalent to detecting 4 ng of superparamagnetic iron oxide (Fe3O4) nanoparticles. The femtoMag was used to quantify the hCG pregnancy hormone by quantifying the number of 200 nm magnetic reporters (superparamagnetic Fe3O4 nanoparticles embedded into a polymer matrix) immuno-captured within the test line of the LFA strip. A sensitivity of 100 pg/mL has been demonstrated. Upon further design and control electronics improvements, the sensitivity is projected to be better than 10 pg/mL. Analysis suggests that an average of 109 hCG molecules are needed to specifically bind 107 nanoparticles in the test line. The ratio of the number of hCG molecules in the sample to the number of reporters in the test line increases monotonically from 20 to 500 as the hCG concentration increases from 0.1 ng/mL to 10 ng/mL. The low-cost easy-to-use femtoMag platform offers high-sensitivity/high-precision target analyte quantification and promises to bring state-of-the-art medical diagnostic tests to the point of care.

scholarly journals Comparative analysis of three point-of-care lateral flow immunoassays for detection of anti-SARS-CoV-2 antibodies in COVID-19 confirmed healthcare workers

2020 ◽  
Author(s):  
Danielle Dias Conte ◽  
Joseane Mayara Almeida Carvalho ◽  
Luciano Kleber de Souza Luna ◽  
Klinger Soares Faíco-Filho ◽  
Ana Helena Perosa ◽  
...  
Keyword(s):  
Healthcare Workers ◽  
Large Scale ◽  
Point Of Care ◽  
Antibody Test ◽  
Cost Effective ◽  
Lateral Flow ◽  
Rt Pcr ◽  
Serum Samples ◽  
Igg Antibody ◽  
Serological Tests

AbstractSince the Coronavirus Disease 2019 (COVID-19) pandemic, Brazil has the third-highest number of confirmed cases and the second-highest number of recovered patients. SARS-CoV-2 detection by real-time RT-PCR is the gold standard in certified infrastructured laboratories. However, for large-scale testing, diagnostics should be fast, cost-effective, widely available, and deployed for the community, such as serological tests based on lateral flow immunoassay (LFIA) for IgM/IgG detection. We evaluated three different commercial point-of-care (POC) LFIAs for anti-SARS-CoV-2 IgM and IgG detection in capillary whole blood of 100 healthcare workers (HCW) previously tested by RT-PCR: 1) COVID-19 IgG/IgM BIO (Bioclin, Brazil), 2) Diagnostic kit for IgM/IgG Antibody to Coronavirus (SARS-CoV-2) (Livzon, China); and 3) SARS-CoV-2 Antibody Test (Wondfo, China). A total of 84 positives and 16 negatives HCW were tested. The data was also analyzed by the number of days post symptoms (DPS) in three groups: <30 (n=26), 30-59 (n=42), and >59 (n=16). Overall detection was 85.71%, 47.62%, and 44.05% for Bioclin, Livzon, and Wondfo, respectively, with a specificity of 100%, and 98.75% for Livzon on storage serum samples. Bioclin was more sensitive (p<0.01), regardless of the DPS. Thus, the Bioclin can be used as a POC test to monitor SARS-CoV-2 seroconversion in HCW.

scholarly journals Duplex Shiny app quantification of the sepsis biomarkers C-reactive protein and interleukin-6 in a fast quantum dot labeled lateral flow assay

2020 ◽  
Vol 18 (1) ◽  
Author(s):  
Christoph Ruppert ◽  
Lars Kaiser ◽  
Lisa Johanna Jacob ◽  
Stefan Laufer ◽  
Matthias Kohl ◽  
...  
Keyword(s):  
Interleukin 6 ◽  
Test Line ◽  
Uv Light ◽  
Point Of Care ◽  
Software Tool ◽  
Lateral Flow ◽  
C Reactive Protein ◽  
Cdse Qds ◽  
Reactive Protein ◽  
Shiny App

Abstract Fast point-of-care (POC) diagnostics represent an unmet medical need and include applications such as lateral flow assays (LFAs) for the diagnosis of sepsis and consequences of cytokine storms and for the treatment of COVID-19 and other systemic, inflammatory events not caused by infection. Because of the complex pathophysiology of sepsis, multiple biomarkers must be analyzed to compensate for the low sensitivity and specificity of single biomarker targets. Conventional LFAs, such as gold nanoparticle dyed assays, are limited to approximately five targets—the maximum number of test lines on an assay. To increase the information obtainable from each test line, we combined green and red emitting quantum dots (QDs) as labels for C-reactive protein (CRP) and interleukin-6 (IL-6) antibodies in an optical duplex immunoassay. CdSe-QDs with sharp and tunable emission bands were used to simultaneously quantify CRP and IL-6 in a single test line, by using a single UV-light source and two suitable emission filters for readout through a widely available BioImager device. For image and data processing, a customized software tool, the MultiFlow-Shiny app was used to accelerate and simplify the readout process. The app software provides advanced tools for image processing, including assisted extraction of line intensities, advanced background correction and an easy workflow for creation and handling of experimental data in quantitative LFAs. The results generated with our MultiFlow-Shiny app were superior to those generated with the popular software ImageJ and resulted in lower detection limits. Our assay is applicable for detecting clinically relevant ranges of both target proteins and therefore may serve as a powerful tool for POC diagnosis of inflammation and infectious events.

scholarly journals Recombinase Polymerase Amplification (RPA) Combined with Lateral Flow Immunoassay for Rapid Detection of Salmonella in Food

Foods ◽  
2019 ◽  
Vol 9 (1) ◽  
pp. 27 ◽  
Author(s):  
Jiali Li ◽  
Biao Ma ◽  
Jiehong Fang ◽  
Antong Zhi ◽  
Erjing Chen ◽  
...  
Keyword(s):  
Test Line ◽  
Point Of Care ◽  
Limited Resource ◽  
Test Strip ◽  
Lateral Flow ◽  
Recombinase Polymerase Amplification ◽  
Lateral Flow Immunoassay ◽  
Quantitative Detection ◽  
Control Line ◽  
Test Strip Reader

Salmonella can cause serious foodborne diseases. We have developed a lateral flow immunoassay combined with recombinase polymerase amplification (LFD-RPA) for detection of Salmonella in food. The conserved fragment (fimY) was selected as the target gene. Under an optimal condition (37 °C, 10 min), the sensitivity was 12 colony-forming units (CFU)/mL in a pure culture. Testing with 16 non-Salmonella strains as controls revealed that LFD-RPA was specific to the fimY gene of Salmonella. The established assay could detect Salmonella at concentrations as low as 1.29 × 102 CFU/mL in artificially contaminated samples. This detection was at a slightly higher level than that for a pure bacterial culture. Combined with the test strip reader, the LFD-RPA is a feasible method for quantitative detection of Salmonella based on the test line intensity, which was the ratio for the test line and control line of the reflected light. The method could be a potential point-of-care test in limited resource areas and provides a new approach and technical support for the diagnosis of food safety.

scholarly journals Pipetting-based immunoassay for point-of-care testing: Application for detection of the influenza A virus

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Ji Yeong Noh ◽  
Sun-Woo Yoon ◽  
Youngji Kim ◽  
Thi Van Lo ◽  
Min-Ju Ahn ◽  
...  
Keyword(s):  
Influenza A Virus ◽  
Influenza A ◽  
Flow System ◽  
Limit Of Detection ◽  
Point Of Care ◽  
Lateral Flow ◽  
Ease Of Use ◽  
Target Antigen ◽  
Enzymatic Reactions

Abstract Point-of-care tests (POCT) for pathogens are considered important for low-resource countries and facilities. Although lateral flow immunoassays (LFIA) have many advantages including speed and ease of use, their sensitivity is limited without specific equipment. Furthermore, their response cannot be enhanced through enzymatic reactions. Owing to these limitations, LFIAs have not yet been generally adopted as the standard protocol for in vitro analysis of infectious pathogens. We aimed to develop a novel pipetting-based immunoassay using a removable magnetic ring-coupled pipette tip. The “magnetic bead-capture antibody-targeted protein complex” was simply purified by pipetting and quantified by enzymatic colour development or using a lateral flow system. This pipetting-based immunoassay was applied to detect the nucleoprotein (NP) of the influenza A virus. Using an HRP-conjugated monoclonal antibody as a probe, the assay allowed for specific and sensitive detection. Furthermore, when this assay was applied exclusively for antigen capture in the lateral flow system, the limit of detection improved 100-fold and displayed greater sensitivity than the lateral flow system alone. Therefore, the pipetting-based immunoassay may be potentially used as a sensitive POCT to clinically detect a target antigen.

scholarly journals Evaluating ELISA, Immunofluorescence, and Lateral Flow Assay for SARS-CoV-2 Serologic Assays

2020 ◽  
Vol 11 ◽  
Author(s):  
Moïse Michel ◽  
Amar Bouam ◽  
Sophie Edouard ◽  
Florence Fenollar ◽  
Fabrizio Di Pinto ◽  
...  
Keyword(s):  
Rapid Test ◽  
Antibody Test ◽  
Immunofluorescence Assay ◽  
Lateral Flow ◽  
Antibody Detection ◽  
Rt Pcr ◽  
Igg Antibody ◽  
Reliable Determination ◽  
Test Kit ◽  
Serological Status

BackgroundThe SARS-CoV-2 outbreak has emerged at the end of 2019. Aside from the detection of viral genome with specific RT-PCR, there is a growing need for reliable determination of the serological status. We aimed at evaluating five SARS-CoV-2 serology assays.MethodsAn in-house immunofluorescence assay (IFA), two ELISA kits (EUROIMMUN® ELISA SARS-CoV-2 IgG and NovaLisa® SARS-CoV-2 IgG and IgM) and two lateral flow assays (T-Tek® SARS-CoV-2 IgG/IgM Antibody Test Kit and Sure Bio-tech® SARS-CoV-2 IgM/IgG Antibody Rapid Test) were compared on 40 serums from RT-PCR-confirmed SARS-CoV-2 infected patients and 10 SARS-CoV-2 RT-PCR negative subjects as controls.ResultsControl subjects tested negative for SARS-CoV-2 antibodies with all five systems. Estimated sensitivities varied from 35.5 to 71.0% for IgG detection and from 19.4 to 64.5% for IgM detection. For IgG, in-house IFA, EuroImmun, T-Tek and NovaLisa displayed 50–72.5% agreement with other systems except IFA vs EuroImmun and T-Tek vs NovaLisa. Intermethod agreement for IgM determination was between 30 and 72.5%.DiscussionThe overall intermethod agreement was moderate. This inconsistency could be explained by the diversity of assay methods, antigens used and immunoglobulin isotype tested. Estimated sensitivities were low, highlighting the limited value of antibody detection in CoVID-19.ConclusionComparison of five systems for SARS-CoV-2 IgG and IgM antibodies showed limited sensitivity and overall concordance. The place and indications of serological status assessment with currently available tools in the CoVID-19 pandemic need further evaluations.

scholarly journals Aptamer-Based Lateral Flow Assays: Current Trends in Clinical Diagnostic Rapid Tests

2022 ◽  
Vol 15 (1) ◽  
pp. 90
Author(s):  
Marjan Majdinasab ◽  
Mihaela Badea ◽  
Jean Louis Marty
Keyword(s):  
Point Of Care ◽  
Good Alternative ◽  
Lateral Flow ◽  
Cold Chain ◽  
High Price ◽  
Clinical Diagnostic ◽  
Lateral Flow Assays ◽  
Rapid Tests ◽  
Detection Of Biomarkers

The lateral flow assay (LFA) is an extensively used paper-based platform for the rapid and on-site detection of different analytes. The method is user-friendly with no need for sophisticated operation and only includes adding sample. Generally, antibodies are employed as the biorecognition elements in the LFA. However, antibodies possess several disadvantages including poor stability, high batch-to-batch variation, long development time, high price and need for ethical approval and cold chain. Because of these limitations, aptamers screened by an in vitro process can be a good alternative to antibodies as biorecognition molecules in the LFA. In recent years, aptamer-based LFAs have been investigated for the detection of different analytes in point-of-care diagnostics. In this review, we summarize the applications of aptamer technology in LFAs in clinical diagnostic rapid tests for the detection of biomarkers, microbial analytes, hormones and antibiotics. Performance, advantages and drawbacks of the developed assays are also discussed.

Ultrastructural Investigations of the Contractile Filaments of Amoebae

1974 ◽  
Vol 32 ◽  
pp. 188-189
Author(s):  
P.L. Moore
Keyword(s):  
Cytoplasmic Streaming ◽  
Three Dimensional ◽  
Freeze Fracture ◽  
Amoeboid Movement ◽  
Different Types ◽  
Chemical Conditions ◽  
Complete Interpretation

Previous freeze fracture results on the intact giant, amoeba Chaos carolinensis indicated the presence of a fibrillar arrangement of filaments within the cytoplasm. A complete interpretation of the three dimensional ultrastructure of these structures, and their possible role in amoeboid movement was not possible, since comparable results could not be obtained with conventional fixation of intact amoebae. Progress in interpreting the freeze fracture images of amoebae required a more thorough understanding of the different types of filaments present in amoebae, and of the ways in which they could be organized while remaining functional.The recent development of a calcium sensitive, demembranated, amoeboid model of Chaos carolinensis has made it possible to achieve a better understanding of such functional arrangements of amoeboid filaments. In these models the motility of demembranated cytoplasm can be controlled in vitro, and the chemical conditions necessary for contractility, and cytoplasmic streaming can be investigated. It is clear from these studies that “fibrils” exist in amoeboid models, and that they are capable of contracting along their length under conditions similar to those which cause contraction in vertebrate muscles.

Correlated Studies of Cellular Interactions Using TEM, Freeze Etching, and SEM

1974 ◽  
Vol 32 ◽  
pp. 320-321
Author(s):  
J. P. Revel
Keyword(s):  
Developmental Stages ◽  
Three Dimensional ◽  
Cell Movement ◽  
Cellular Interactions ◽  
Serial Sections ◽  
Cell Sheets ◽  
Freeze Etching ◽  
Early Developmental

Movement of individual cells or of cell sheets and complex patterns of folding play a prominent role in the early developmental stages of the embryo. Our understanding of these processes is based on three- dimensional reconstructions laboriously prepared from serial sections, and from autoradiographic and other studies. Many concepts have also evolved from extrapolation of investigations of cell movement carried out in vitro. The scanning electron microscope now allows us to examine some of these events in situ. It is possible to prepare dissections of embryos and even of tissues of adult animals which reveal existing relationships between various structures more readily than used to be possible vithout an SEM.

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