scholarly journals Rapid Multiplexed Detection on Lateral-Flow Devices Using a Laser Direct-Write Technique

Biosensors ◽  
2018 ◽  
Vol 8 (4) ◽  
pp. 97 ◽  
Author(s):  
Peijun He ◽  
Ioannis Katis ◽  
Robert Eason ◽  
Collin Sones
Keyword(s):  
Bacterial Infections ◽  
Point Of Care ◽  
Simultaneous Detection ◽  
Cross Reactivity ◽  
Lateral Flow ◽  
Direct Write ◽  
Multiplexed Detection ◽  
Amyloid A ◽  
Flow Devices ◽  
Laser Direct Write

Paper-based lateral flow devices (LFDs) are regarded as ideal low-cost diagnostic solutions for point-of-care (POC) scenarios that allow rapid detection of a single analyte within a fluidic sample, and have been in common use for a decade. In recent years, there has been an increasing need for rapid and simultaneous detection of multiple analytes present within a single sample and to facilitate this, we report here a novel solution—detection using a multi-path LFD created via the precise partitioning of the single flow-path of a standard LFD using our previously reported laser direct-write (LDW) technique. The multiple flow-paths allow the simultaneous detection of the different analytes individually within each of the parallel channels without any cross-reactivity. The appearance of coloured test lines in individual channels indicates the presence of the different analytes within a sample. We successfully present the use of a LDW-patterned multi-path LFD for multiplexed detection of a biomarker panel comprising C-reactive protein (CRP) and Serum amyloid A-1 (SAA1), used for the diagnosis of bacterial infections. Overall, we demonstrate the use of our LDW technique in the creation of a novel LFD that enables multiplexed detection of two inflammation markers within a single LFD providing a detection protocol that is comparatively more efficient than the standard sequential multiplexing procedure.

scholarly journals A rapid diagnostic test for human Visceral Leishmaniasis using novel Leishmania antigens in a Laser Direct-Write Lateral Flow Device

2019 ◽  
Vol 8 (1) ◽  
pp. 1178-1185 ◽  
Author(s):  
Maria Victoria Humbert ◽  
Lourena Emanuele Costa ◽  
Ioannis Katis ◽  
Fernanda Fonseca Ramos ◽  
Amanda Sanchéz Machado ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Diagnostic Test ◽  
Rapid Diagnostic Test ◽  
Lateral Flow ◽  
Direct Write ◽  
Lateral Flow Device ◽  
Human Visceral Leishmaniasis ◽  
Flow Device ◽  
Laser Direct Write

Multiplexed detection of biomarkers in lateral-flow immunoassays

The Analyst ◽  
2020 ◽  
Vol 145 (8) ◽  
pp. 2828-2840 ◽  
Author(s):  
Lei Huang ◽  
Shulin Tian ◽  
Wenhao Zhao ◽  
Ke Liu ◽  
Xing Ma ◽  
...  
Keyword(s):  
Simultaneous Detection ◽  
Lateral Flow ◽  
Diagnostic Efficiency ◽  
Multiplexed Detection ◽  
Multiple Biomarkers ◽  
Diagnostic Cost ◽  
Detection Of Biomarkers

Multiplexed detection of biomarkers, i.e., simultaneous detection of multiple biomarkers in a single assay, can enhance diagnostic precision, improve diagnostic efficiency, reduce diagnostic cost, and alleviate pain of patients.

scholarly journals Multiple Bacteria Identification in the Point-of-Care: an Old Method Serving a New Approach

Sensors ◽  
2020 ◽  
Vol 20 (12) ◽  
pp. 3351
Author(s):  
Sara Viveiros ◽  
Mónica Rodrigues ◽  
Débora Albuquerque ◽  
Sofia A. M. Martins ◽  
Susana Cardoso ◽  
...  
Keyword(s):  
Bacterial Infections ◽  
Limit Of Detection ◽  
Detection System ◽  
Point Of Care ◽  
Bovine Mastitis ◽  
Cross Reactivity ◽  
Superparamagnetic Nanoparticles ◽  
Nucleic Acid Amplification ◽  
Streptococcus Uberis ◽  
Detection Assay

The accurate diagnosis of bacterial infections is of critical importance for effective treatment decisions. Due to the multietiologic nature of most infectious diseases, multiplex assays are essential for diagnostics. However, multiplexability in nucleic acid amplification-based methods commonly resorts to multiple primers and/or multiple reaction chambers, which increases analysis cost and complexity. Herein, a polymerase chain reaction (PCR) offer method based on a universal pair of primers and an array of specific oligonucleotide probes was developed through the analysis of the bacterial 16S ribosomal RNA gene. The detection system consisted of DNA hybridization over an array of magnetoresistive sensors in a microfabricated biochip coupled to an electronic reader. Immobilized probes interrogated single-stranded biotinylated amplicons and were obtained using asymmetric PCR. Moreover, they were magnetically labelled with streptavidin-coated superparamagnetic nanoparticles. The benchmarking of the system was demonstrated to detect five major bovine mastitis-causing pathogens: Escherichia coli, Klebsiella sp., Staphylococcus aureus, Streptococcus uberis, and Streptococcus agalactiae. All selected probes proved to specifically detect their respective amplicon without significant cross reactivity. A calibration curve was performed for S. agalactiae, which demonstrates demonstrating a limit of detection below 30 fg/µL. Thus, a sensitive and specific multiplex detection assay was established, demonstrating its potential as a bioanalytical device for point-of-care applications.

scholarly journals Evaluation of Six Commercial Mid- to High-Volume Antibody and Six Point-of-Care Lateral Flow Assays for Detection of SARS-CoV-2 Antibodies

2020 ◽  
Vol 58 (10) ◽  
Author(s):  
Carmen L. Charlton ◽  
Jamil N. Kanji ◽  
Kam Johal ◽  
Ashley Bailey ◽  
Sabrina S. Plitt ◽  
...  
Keyword(s):  
Symptom Onset ◽  
Point Of Care ◽  
Acute Infection ◽  
High Volume ◽  
Respiratory Viruses ◽  
Cross Reactivity ◽  
Lateral Flow ◽  
Serological Tests ◽  
Deep Blue ◽  
Lateral Flow Assays

ABSTRACT Coronavirus disease (COVID) serological tests are essential to determine the overall seroprevalence of a population and to facilitate exposure estimates within that population. We performed a head-to-head assessment of enzyme immunoassays (EIAs) and point-of-care lateral flow assays (POCTs) to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibodies. Demographics, symptoms, comorbidities, treatment, and mortality of patients whose sera were used were also reviewed. Six EIAs (Abbott, Affinity, Bio-Rad, DiaSorin, Euroimmun, and Roche) and six POCTs (BTNX, Biolidics, Deep Blue, Genrui, Getein BioTech, and Innovita) were evaluated for the detection of SARS-CoV-2 antibodies in known COVID-19-infected individuals. Sensitivity of EIAs ranged from 50 to 100%, with only four assays having overall sensitivities of >95% after 21 days after symptom onset. Notably, cross-reactivity with other respiratory viruses (parainfluenza virus [PIV-4] [n = 5], human metapneumovirus [hMPV] [n = 3], rhinovirus/enterovirus [n = 1], CoV-229E [n = 2], CoV-NL63 [n = 2], and CoV-OC43 [n = 2]) was observed; however, overall specificity of EIAs was good (92 to 100%; all but one assay had specificity above 95%). POCTs were 0 to 100% sensitive >21 days after onset, with specificity ranging from 96 to 100%. However, many POCTs had faint banding and were often difficult to interpret. Serology assays can detect SARS-CoV-2 antibodies as early as 10 days after symptom onset. Serology assays vary in their sensitivity based on the marker (IgA/IgM versus IgG versus total) and by manufacturer; however, overall only 4 EIAs and 4 POCTs had sensitivities of >95% >21 days after symptom onset. Cross-reactivity with other seasonal coronaviruses is of concern. Serology assays should not be used for the diagnosis of acute infection but rather in carefully designed serosurveys to facilitate understanding of seroprevalence in a population and to identify previous exposure to SARS-CoV-2.

scholarly journals A nanofluidic device for rapid and multiplexed SARS-CoV-2 serological antibody detection

2021 ◽  
Author(s):  
Thomas Mortelmans ◽  
Dimitrios Kazazis ◽  
Celestino Padeste ◽  
Philipp Berger ◽  
Xiaodan Li ◽  
...  
Keyword(s):  
Influenza A ◽  
Point Of Care ◽  
Simultaneous Detection ◽  
High Sensitivity ◽  
Modern Society ◽  
Antibody Detection ◽  
Multiplexed Detection ◽  
Viral Spread ◽  
Point Of Care Diagnostics ◽  
Nanofluidic Device

Abstract The outbreak of COVID-19 has led to a substantial death toll and has hindered the functioning of modern society, sending the world into a medical and economic crisis1,2. This underlined the importance of point-of-care diagnostics, as well as accurate, cost-effective serological antibody tests as well as point-of-care diagnostics to monitor the viral spread and contain pandemics and endemics. Here, we present a three-dimensional (3D) nanofluidic device for rapid and multiplexed detection of viral antibodies. The device is designed to size-dependently immobilize particles from a multi-particle mixture at predefined positions in nanochannels through capillary forces only, resulting in distinct trapping lines. We show that individual lines can be used as an on-chip fluorescence-linked immunosorbent assay for multiplexed detection of serological immunoglobulin antibodies against viral proteins with high sensitivity. Further device versatility is exhibited by on-bead color multiplexing for simultaneous detection of IgG and IgM antibodies in convalescent human serum and by concurrent detection of anti-spike (SARS-CoV-2) and anti-hemagglutinin (Influenza A) antibodies. The device’s applications can be further extended to detect a plethora of diseases simultaneously in a reliable and straightforward manner.

scholarly journals Upconverting phosphor technology-based lateral flow assay for the rapid and sensitive detection of anti-Trichinella spiralis IgG antibodies in pig serum

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Jian Li ◽  
Jing Ding ◽  
Xiao-Lei Liu ◽  
Bin Tang ◽  
Xue Bai ◽  
...  
Keyword(s):  
Trichinella Spiralis ◽  
Point Of Care ◽  
Cross Reactivity ◽  
Lateral Flow ◽  
Sensitive Detection ◽  
Parasitic Infections ◽  
Igg Antibodies ◽  
Point Of Care Test ◽  
Rabbit Igg ◽  
Pig Serum

Abstract Background Trichinella spiralis is a zoonotic food-borne parasite. A disease caused by infection with T. spiralis is called trichinellosis in humans. It is important to investigate the epidemic situation and the surveillance of herds and then prevent infection in humans. Therefore, this study is to develop a rapid and sensitive diagnostic method for on-site test in domestic and wild animals. Methods Upconverting phosphor nanoparticles (UCNPs), an excellent optical label, were conjugated with the excretory-secretory (ES) antigens from T. spiralis muscle larvae (ML) or goat anti-rabbit IgG, and a lateral flow (LF) assay based on these probes (UCNPs-ES/goat anti-rabbit IgG) was developed for the rapid and sensitive detection of anti-T. spiralis IgG antibodies in pig serum. The assay is named the UPT-LF-ES assay. In addition, the probes were characterized, and the assay was optimized. A cut-off threshold of the assay was also identified by using 169 known negative pig samples. Performance of the assay to T. spiralis with different infective numbers, cross-reactivity with other parasitic infections, the single-blinded experiment, and coincidence were evaluated with the assay. Results The UPT-LF-ES assay was successfully constructed and optimized based on the probes of UCNPs-ES/goat anti-rabbit IgG. In the pigs infected with 100, 1000, and 10,000 ML, positive results were first presented at 35 days post-infection (dpi), 30 dpi, and 25 dpi, respectively. The assay had no cross-reaction with other parasitic infections. A single-blinded experiment indicated that the sensitivity and specificity of the UPT-LF-ES assay were 100% and 100%, respectively, the area under the receiver operating characteristic (ROC) curve was 1.000. In addition, the value detected by the UPT-LF-ES assay was significantly different between positive and negative samples. Moreover, compared with the “gold standard” magnetic stirrer method, the coincidence rate of the UPT-LF-ES assay was 87.27%, and the kappa (K) coefficient was 0.7454, showing a substantial agreement. Conclusions The UPT-LF-ES assay is a useful point-of-care test (POCT) with T. spiralis in the detection of pig, which contributes to preventing human trichinellosis. Graphical Abstract

scholarly journals Utility of FebriDx in early identification of possible COVID19 infection

2020 ◽  
Author(s):  
Nawazish Karim ◽  
Muhammad Zubair Ashraf ◽  
Muhammad Naeem ◽  
Tahir Anwar ◽  
Hnin Aung ◽  
...  
Keyword(s):  
Respiratory Tract ◽  
Bacterial Infections ◽  
Respiratory Tract Infections ◽  
Point Of Care ◽  
Protein A ◽  
Simultaneous Detection ◽  
Clinical Manifestations ◽  
Molecular Testing ◽  
Delay In Diagnosis ◽  
Tract Infections

Abstract Background Reliable differentiation between uncomplicated and self-limiting acute respiratory tract infections (ARIs) and more severe bacterial respiratory tract infections remains challenging, due to the non-specific clinical manifestations in both systemic bacterial or viral infections. The current COVID-19 pandemic is putting extraordinary strain on healthcare resources. To date, molecular testing is available but has a long turnaround time and therefore cannot provide results at the point-of-care, leading to a delay in results thereby exposing patients to cross-infection and delay in diagnosis (1-3). Methods We prospectively evaluated the utility of FebriDx®, a point-of-care fingerstick blood test that can differentiate viral from bacterial ARIs through simultaneous detection of both Myxovirus-resistance protein A (MxA) and C-reactive protein (CRP), in rapidly determining viral cases requiring immediate isolation and confirmatory molecular testing, from non-infectious patients or bacterial infections that require antibacterial therapy.Results 75 consecutive patients were assessed and 48 eligible cases were tested with FebriDx®. Overall, 35 patients had FebriDx® test viral positive. All 35 patients had either positive rt-PCR (n=30) for COVID-19 or clinical picture highly suggestive of COVID-19 infection (PPV of 100% in a pandemic situation)[AB1] . In the 13 cases it was viral negative, rRT-PCR was also negative in all cases. In one case of LRTI, it was not possible to determine the exact cause of infection and a viral infection couldn’t be excluded. Including this patient, the NPV was 12/13 (92%) exceeding the NPV of rRt-PCR at 71% (12/17). Sensitivity was conservatively calculated at 97% (35/36) compared to 85.7% (30[RS2] /35) for rRt-PCR. Similarly the specificity of both FebriDx®and rRt-PCR was 100% (12/12).Conclusions In the current COVID-19, FebriDx® shows potential as a reliable POC test and a proxy marker of COVID-19 infection amongst inpatients in a secondary care setting. [AB1]35/35 equates to a sensitivity and specificity of 100% for COVID, would you be willing to say that instead of ‘near 100% ppv)? [RS2]I believe PCR was 85.7% (30/35), because PCR only detects the COVID cases

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