scholarly journals System Architecture for IIoT-Based POC Molecular Diagnostic Device

2021 ◽  
Vol 6 (1) ◽  
pp. 60
Author(s):  
Byeong-Heon Kil ◽  
Ji-Seong Park ◽  
Chan-Young Park ◽  
Yu-Seop Kim ◽  
Jong-Dae Kim
Keyword(s):  
Point Of Care ◽  
Low Cost ◽  
Diagnostic System ◽  
Pcr Amplification ◽  
Dna Amplification ◽  
System Structure ◽  
World Health ◽  
Molecular Diagnostic ◽  
Target System ◽  
Open Platform

In this paper, we investigate an efficient structure for a point-of-care (POC) molecular diagnostic system based on the industrial Internet of things (IIoT). The target system can perform automated molecular diagnosis including DNA extraction, PCR amplification, and fluorescence detection. Samples and reagents are placed in a multi-room cartridge and loaded into the system. A rotating motor and a syringe motor control the cartridge to extract DNA from the sample. The extracted DNA is transferred to a polymerase chain reaction (PCR) chamber for DNA amplification and detection. The proposed system provides multiplexing of up to four colors. For POC molecular diagnostics, the World Health Organization demands features such as low volume, low cost, fast results, and a user-friendly interface. In this paper, we propose a system structure that can satisfy these requirements by using a PCR chip and open platform. A distributed structure is adopted for the convenience of maintenance, and a web-based GUI is adopted for the user’s convenience. We also investigated communication problems that may occur between system components. Using the proposed structure, the user can conveniently control from standard computing devices including a smartphone.

scholarly journals Detection of ESKAPE Bacterial Pathogens at the Point of Care Using Isothermal DNA-Based Assays in a Portable Degas-Actuated Microfluidic Diagnostic Assay Platform

2016 ◽  
Vol 83 (4) ◽  
Author(s):  
Lars D. Renner ◽  
Jindong Zan ◽  
Linda I. Hu ◽  
Manuel Martinez ◽  
Pedro J. Resto ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Point Of Care ◽  
Low Cost ◽  
Bacterial Pathogens ◽  
Diagnostic System ◽  
Molecular Diagnostic ◽  
Technology Gap ◽  
Content Type ◽  
Antimicrobial Drug Resistance ◽  
Industrial Processing

ABSTRACT An estimated 1.5 billion microbial infections occur globally each year and result in ∼4.6 million deaths. A technology gap associated with commercially available diagnostic tests in remote and underdeveloped regions prevents timely pathogen identification for effective antibiotic chemotherapies for infected patients. The result is a trial-and-error approach that is limited in effectiveness, increases risk for patients while contributing to antimicrobial drug resistance, and reduces the lifetime of antibiotics. This paper addresses this important diagnostic technology gap by describing a low-cost, portable, rapid, and easy-to-use microfluidic cartridge-based system for detecting the ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp.) bacterial pathogens that are most commonly associated with antibiotic resistance. The point-of-care molecular diagnostic system consists of a vacuum-degassed microfluidic cartridge preloaded with lyophilized recombinase polymerase amplification (RPA) assays and a small portable battery-powered electronic incubator/reader. The isothermal RPA assays detect the targeted ESKAPE pathogens with high sensitivity (e.g., a limit of detection of ∼10 nucleic acid molecules) that is comparable to that of current PCR-based assays, and they offer advantages in power consumption, engineering, and robustness, which are three critical elements required for the point-of-care setting. IMPORTANCE This paper describes a portable system for rapidly identifying bacteria in resource-limited environments; we highlight the capabilities of the technology by detecting different pathogens within the ESKAPE collection, which cause nosocomial infections. The system is designed around isothermal DNA-based assays housed within an autonomous plastic cartridge that are designed with the end user in mind, who may have limited technological training. Displaying excellent sensitivity and specificity, the assay systems that we demonstrate may enable future diagnoses of bacterial infection to guide the development of effective chemotherapies and may have a role in areas beyond health where rapid detection is valuable, including in industrial processing and manufacturing, food security, agriculture, and water quality testing.

scholarly journals Validation of Differentially Expressed Immune Biomarkers in Latent and Active Tuberculosis by Real-Time PCR

2021 ◽  
Vol 11 ◽  
Author(s):  
Prem Perumal ◽  
Mohamed Bilal Abdullatif ◽  
Harriet N. Garlant ◽  
Isobella Honeyborne ◽  
Marc Lipman ◽  
...  
Keyword(s):  
Low Cost ◽  
Peripheral Blood Leukocyte ◽  
Primary Diagnosis ◽  
Diagnostic Methods ◽  
World Health ◽  
Molecular Diagnostic ◽  
Roc Curve Analysis ◽  
Simple Arithmetic ◽  
Immune Biomarkers ◽  
The Uk

Tuberculosis (TB) remains a major global threat and diagnosis of active TB ((ATB) both extra-pulmonary (EPTB), pulmonary (PTB)) and latent TB (LTBI) infection remains challenging, particularly in high-burden countries which still rely heavily on conventional methods. Although molecular diagnostic methods are available, e.g., Cepheid GeneXpert, they are not universally available in all high TB burden countries. There is intense focus on immune biomarkers for use in TB diagnosis, which could provide alternative low-cost, rapid diagnostic solutions. In our previous gene expression studies, we identified peripheral blood leukocyte (PBL) mRNA biomarkers in a non-human primate TB aerosol-challenge model. Here, we describe a study to further validate select mRNA biomarkers from this prior study in new cohorts of patients and controls, as a prerequisite for further development. Whole blood mRNA was purified from ATB patients recruited in the UK and India, LTBI and two groups of controls from the UK (i) a low TB incidence region (CNTRLA) and (ii) individuals variably-domiciled in the UK and Asia ((CNTRLB), the latter TB high incidence regions). Seventy-two mRNA biomarker gene targets were analyzed by qPCR using the Roche Lightcycler 480 qPCR platform and data analyzed using GeneSpring™ 14.9 bioinformatics software. Differential expression of fifty-three biomarkers was confirmed between MTB infected, LTBI groups and controls, seventeen of which were significant using analysis of variance (ANOVA): CALCOCO2, CD52, GBP1, GBP2, GBP5, HLA-B, IFIT3, IFITM3, IRF1, LOC400759 (GBP1P1), NCF1C, PF4V1, SAMD9L, S100A11, TAF10, TAPBP, and TRIM25. These were analyzed using receiver operating characteristic (ROC) curve analysis. Single biomarkers and biomarker combinations were further assessed using simple arithmetic algorithms. Minimal combination biomarker panels were delineated for primary diagnosis of ATB (both PTB and EPTB), LTBI and identifying LTBI individuals at high risk of progression which showed good performance characteristics. These were assessed for suitability for progression against the standards for new TB diagnostic tests delineated in the published World Health Organization (WHO) technology product profiles (TPPs).

scholarly journals Loop-Mediated Isothermal Amplification as Point-of-Care Diagnosis for Neglected Parasitic Infections

2020 ◽  
Vol 21 (21) ◽  
pp. 7981
Author(s):  
Catalina Avendaño ◽  
Manuel Alfonso Patarroyo
Keyword(s):  
Neglected Tropical Diseases ◽  
Point Of Care ◽  
Low Cost ◽  
High Specificity ◽  
World Health Organisation ◽  
Isothermal Amplification ◽  
World Health ◽  
Parasitic Infections ◽  
River Blindness ◽  
Loop Mediated Isothermal Amplification

The World Health Organisation (WHO) has placed twenty diseases into a group known as neglected tropical diseases (NTDs), twelve of them being parasitic diseases: Chagas’ disease, cysticercosis/taeniasis, echinococcosis, food-borne trematodiasis, human African trypanosomiasis (sleeping sickness), leishmaniasis, lymphatic filariasis, onchocerciasis (river blindness), schistosomiasis, soil-transmitted helminthiasis (ascariasis, hookworm, trichuriasis), guinea-worm and scabies. Such diseases affect millions of people in developing countries where one of the main problems concerning the control of these diseases is diagnosis-based due to the most affected areas usually being far from laboratories having suitable infrastructure and/or being equipped with sophisticated equipment. Advances have been made during the last two decades regarding standardising and introducing techniques enabling diagnoses to be made in remote places, i.e., the loop-mediated isothermal amplification (LAMP) technique. This technique’s advantages include being able to perform it using simple equipment, diagnosis made directly in the field, low cost of each test and the technique’s high specificity. Using this technique could thus contribute toward neglected parasite infection (NPI) control and eradication programmes. This review describes the advances made to date regarding LAMP tests, as it has been found that even though several studies have been conducted concerning most NPI, information is scarce for others.

scholarly journals Highly performing point-of-care molecular testing for SARS-CoV-2 with RNA extraction and isothermal amplification.

2020 ◽  
Author(s):  
Pierre Garneret ◽  
Etienne Coz ◽  
Elian Martin ◽  
Jean-Claude Manuguerra ◽  
Elodie Brient-Litzler ◽  
...  
Keyword(s):  
Point Of Care ◽  
Low Cost ◽  
Rna Extraction ◽  
Dna Amplification ◽  
Developed Countries ◽  
Temperature Cycling ◽  
Isothermal Amplification ◽  
Nucleic Acid Amplification ◽  
Clinical Samples ◽  
Molecular Testing

In order to respond to the urgent request of massive testing, developed countries perform nucleic acid amplification tests (NAAT) of SARS-CoV-2 in centralized laboratories. Real-time RT - PCR (Reverse transcription - Polymerase Chain Reaction) is used to amplify the viral RNA and enable its detection. Although PCR is 37 years old, it is still considered, without dispute, as the gold standard. PCR is an efficient process, but the complex engineering required for automated RNA extraction and temperature cycling makes it incompatible for use in point of care settings. In the present work, by harnessing progress made in the past two decades in DNA amplification, microfluidics and membrane technologies, we succeeded to create a portable test, in which SARS-CoV-2 RNA is extracted, amplified isothermally by RT - LAMP (Loop-mediated Isothermal Amplification), and detected using intercalating dyes or highly fluorescent probes. Depending on the viral load, the detection takes between twenty minutes and one hour. Using pools of naso-pharyngal clinical samples, we estimated a sensitivity comparable to RT-qPCR (up to a Cycle threshold of 39, equivalent to <0.1 TCID50 per mL) and a 100% specificity, for other human coronaviruses and eight respiratory viruses currently circulating in Europe. We designed and fabricated an easy-to-use portable device called COVIDISC to carry out the test at the point of care. The low cost of the materials along with the absence of complex equipment paves the way towards a large dissemination of this device. The perspective of a reliable SARS-CoV-2 point of care detection, highly performing, that would deliver on-site results in less than one hour opens up a new efficient approach to manage the pandemics.

scholarly journals A fully integrated paperfluidic molecular diagnostic chip for the extraction, amplification, and detection of nucleic acids from clinical samples

Lab on a Chip ◽  
2016 ◽  
Vol 16 (4) ◽  
pp. 753-763 ◽  
Author(s):  
Natalia M. Rodriguez ◽  
Winnie S. Wong ◽  
Lena Liu ◽  
Rajan Dewar ◽  
Catherine M. Klapperich
Keyword(s):  
Nucleic Acids ◽  
Point Of Care ◽  
Low Cost ◽  
Molecular Diagnostic ◽  
Clinical Samples ◽  
Fully Integrated

We present a low-cost, disposable, and fully-integrated paperfluidic molecular diagnostic chip for sample-to-result functionality at the point-of-care.

scholarly journals SARS-CoV-2 detection with de novo designed synthetic riboregulators

2020 ◽  
Author(s):  
Ilkay Cisil Koksaldi ◽  
Recep Erdem Ahan ◽  
Sila Kose ◽  
Nedim Haciosmanoglu ◽  
Ebru Sahin Kehribar ◽  
...  
Keyword(s):  
De Novo ◽  
Point Of Care ◽  
Low Cost ◽  
Critical Role ◽  
World Health ◽  
Global Pandemic ◽  
Health Organization ◽  
Diagnostic Capabilities ◽  
Synthetic Biology Approach

Sars-CoV-2 is a human pathogen and is the main cause of COVID-19 disease. COVID-19 is announced as a global pandemic by World Health Organization. COVID-19 is characterized by severe conditions and early diagnosis can make dramatic changes both for personal and public health. In order to increase the reach for low cost equipment which requires a very limited technical knowledge can be beneficial to diagnose the viral infection. Such diagnostic capabilities can have a very critical role to control the transmission of the disease. Here we are reporting a state-of-the-art diagnostic tool developed by using an in vitro synthetic biology approach by employing engineered de novo riboregulators. Our design coupled with a home-made point-of-care device setting can detect and report presence of Sars-CoV-2 specific genes. The presence of Sars-CoV-2 related genes triggers translation of sfGFP mRNAs, resulting in green fluorescence output. The approach proposed here has the potential of being a game changer in Sars-COV-2 diagnostics by providing an easy-to-run, low-cost-demanding diagnostic capability.

scholarly journals Design and Experimental Investigation of a Novel Spiral Microfluidic Chip to Separate Wide Size Range of Micro-Particles Aimed at Cell Separation

2021 ◽  
Author(s):  
Seyed Ali Tabatabaei ◽  
Mohammad Zabetian targhi
Keyword(s):  
Cell Separation ◽  
Point Of Care ◽  
Low Cost ◽  
Diagnostic System ◽  
Average Diameter ◽  
Microfluidic System ◽  
Microfluidic Chips ◽  
Blood Samples ◽  
Micro Particles ◽  
User Friendly

Abstract BackgroundIsolation of microparticles and biological cells on microfluidic chips has received considerable attention due to their applications in numerous areas such as medical and engineering fields. Microparticles separation are of great importance in bioassays owing to the need for a smaller sample and device size, and lower manufacturing costs. In this study, we first explain the concepts of separation and microfluidic science along with their applications in the medical sciences, and then, a conceptual design of a novel inertial microfluidic system is proposed and analyzed. The PDMS spiral microfluidic device was fabricated, and its effects on the separation of particles with sizes similar to biological particles were experimentally analyzed. This separation technique can be used in the process of separating cancer cells from the normal ones in the blood samples.ResultsThese components required for testing were selected, assembled, and finally, a very affordable microfluidic kit was provided. Different experiments were designed, and the results were analyzed using appropriate software and methods. Separator system tests with polydisperse hollow glass particles (diameter 2-20 µm), and monodisperse Polystyrene particles (diameter 5,15 µm), and the results exhibit an acceptable chip performance with 86 percent of efficiency for both monodisperse particles and polydisperse particles. The microchannel collects particles with an average diameter of 15.8 μm, 9.4 μm, and 5.9 μm at the Proposed reservoirs. ConclusionThis chip can be integrated into a more extensive point-of-care diagnostic system to test blood samples, and it could be said Based on the results of the experiments, this low-cost and user-friendly setting can be used for a variety of microparticle separation programs such as cell separation in biological assays.

scholarly journals Bioluminescent detection of isothermal DNA amplification in microfluidic generated droplets and artificial cells

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Patrick Hardinge ◽  
Divesh K. Baxani ◽  
Thomas McCloy ◽  
James A. H. Murray ◽  
Oliver K. Castell
Keyword(s):  
Real Time ◽  
High Throughput ◽  
Soft Matter ◽  
Low Cost ◽  
Diagnostic Technique ◽  
Dna Amplification ◽  
Droplet Microfluidics ◽  
Nucleic Acid Amplification ◽  
Molecular Diagnostic ◽  
Artificial Cells

AbstractMicrofluidic droplet generation affords precise, low volume, high throughput opportunities for molecular diagnostics. Isothermal DNA amplification with bioluminescent detection is a fast, low-cost, highly specific molecular diagnostic technique that is triggerable by temperature. Combining loop-mediated isothermal nucleic acid amplification (LAMP) and bioluminescent assay in real time (BART), with droplet microfluidics, should enable high-throughput, low copy, sequence-specific DNA detection by simple light emission. Stable, uniform LAMP–BART droplets are generated with low cost equipment. The composition and scale of these droplets are controllable and the bioluminescent output during DNA amplification can be imaged and quantified. Furthermore these droplets are readily incorporated into encapsulated droplet interface bilayers (eDIBs), or artificial cells, and the bioluminescence tracked in real time for accurate quantification off chip. Microfluidic LAMP–BART droplets with high stability and uniformity of scale coupled with high throughput and low cost generation are suited to digital DNA quantification at low template concentrations and volumes, where multiple measurement partitions are required. The triggerable reaction in the core of eDIBs can be used to study the interrelationship of the droplets with the environment and also used for more complex chemical processing via a self-contained network of droplets, paving the way for smart soft-matter diagnostics.

scholarly journals A Complete Protocol for Rapid and Low-Cost Fabrication of Polymer Microfluidic Chips Containing Three-Dimensional Microstructures Used in Point-of-Care Devices

Micromachines ◽  
2019 ◽  
Vol 10 (9) ◽  
pp. 624 ◽  
Author(s):  
Trieu Nguyen ◽  
Aaydha Chidambara Vinayaka ◽  
Dang Duong Bang ◽  
Anders Wolff
Keyword(s):  
Salmonella Enteritidis ◽  
Solid Phase ◽  
Point Of Care ◽  
Low Cost ◽  
Three Dimensional ◽  
Diagnostic Methods ◽  
Molecular Diagnostic ◽  
Microfluidic Chips ◽  
Industrial Companies ◽  
Polymerase Chain

This protocol provides insights into the rapid, low-cost, and largescale fabrication of polymer microfluidic chips containing three-dimensional microstructures used in point-of-care devices for applications such as detection of pathogens via molecular diagnostic methods. The details of the fabrication methods are described in this paper. This study offers suggestions for researchers and experimentalists, both at university laboratories and in industrial companies, to prevent doom fabrication issues. For a demonstration of bio-application in point-of-care testing, the 3D microarrays fabricated are then employed in multiplexed detection of Salmonella (Salmonella Typhimurium and Salmonella Enteritidis), based on a molecular detection technique called solid-phase polymerase chain reaction (SP-PCR).

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