scholarly journals A robust, hand-powered, instrument-free sample preparation system for point-of-care pathogen detection

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Fei Zhao ◽  
Eun Yeong Lee ◽  
Geun Su Noh ◽  
Jaehyup Shin ◽  
Huifang Liu ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Sample Preparation ◽  
Pathogen Detection ◽  
Point Of Care ◽  
Low Cost ◽  
Sample Pretreatment ◽  
Sample Volume ◽  
Nucleic Acid Isolation ◽  
Point Of Care Diagnostics ◽  
Reaction Matrix

Abstract Here, we describe a simple, universal protocol for use in nucleic acid testing-based pathogen diagnostics, which requires only hand-powered sample preparation, including the processes of pathogen enrichment and nucleic acid isolation. The protocol uses low-cost amine-functionalized diatomaceous earth with a 1-μm Teflon filter as a reaction matrix in both stages of the process, using homobifunctional imidoesters. Using a simple syringe as a pump, the capture efficiency for a large sample volume (<50 mL) was enhanced by up to 98.3%, and the detection limit was 1 CFU/mL, 100-fold better than that of common commercial nucleic acid isolation kit. This protocol can also be combined with commercialized 96-well filter plates for robust sample preparation. Our proposed system is robust, simple, low-cost, universal, and rapid (taking <20 min), and it works regardless of the ambient environment and sample pretreatment, requiring no electricity or instruments. Its benefits include the simplicity of producing its components and its ease of operation, and it can be readily integrated with other assays for point-of-care diagnostics.

scholarly journals A Robust, Low-Cost Instrument for Real-time Colorimetric Isothermal Nucleic Acid Amplification

2021 ◽  
Author(s):  
Frank Myers ◽  
Brian Moffatt ◽  
Ragheb El Khaja ◽  
Titash Chatterjee ◽  
Gurmeet Marwaha ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Real Time ◽  
Point Of Care ◽  
Low Cost ◽  
Lamp Assay ◽  
Nucleic Acid Amplification ◽  
Diagnostic Assays ◽  
Isothermal Nucleic Acid Amplification ◽  
Point Of Care Diagnostics ◽  
Time Readout

The COVID-19 pandemic has highlighted the need for broader access to molecular diagnostics. Colorimetric isothermal nucleic acid amplification assays enable simplified instrumentation over more conventional PCR diagnostic assays and, as such, represent a promising approach for addressing this need. In particular, colorimetric LAMP (loop-mediated isothermal amplification) has received a great deal of interest recently. However, there do not currently exist robust instruments for performing these kinds of assays in high throughput with real-time readout of amplification signals. To address this need, we developed LARI, the LAMP Assay Reader Instrument. We have deployed over 50 LARIs for routine use in R&D and production environments, with over 12,000 assays run to date. In this paper, we present the design and construction of LARI along with thermal, optical, and assay performance characteristics. LARI can be produced for under $1500 and has broad applications in R&D, point-of-care diagnostics, and global health.

Paper-based nucleic acid sample preparation for point-of-care diagnostics

2021 ◽  
pp. 131272
Author(s):  
Shruti Soni ◽  
Bhushan J. Toley
Keyword(s):  
Nucleic Acid ◽  
Sample Preparation ◽  
Point Of Care ◽  
Point Of Care Diagnostics ◽  
Acid Sample

scholarly journals A glucose meter interface for point-of-care gene circuit-based diagnostics

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Evan Amalfitano ◽  
Margot Karlikow ◽  
Masoud Norouzi ◽  
Katariina Jaenes ◽  
Seray Cicek ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Synthetic Biology ◽  
Molecular Diagnostics ◽  
Point Of Care ◽  
Low Cost ◽  
Gene Circuit ◽  
Reporter Systems ◽  
Glucose Meter ◽  
Pandemic Response ◽  
Glucose Output

AbstractRecent advances in cell-free synthetic biology have given rise to gene circuit-based sensors with the potential to provide decentralized and low-cost molecular diagnostics. However, it remains a challenge to deliver this sensing capacity into the hands of users in a practical manner. Here, we leverage the glucose meter, one of the most widely available point-of-care sensing devices, to serve as a universal reader for these decentralized diagnostics. We describe a molecular translator that can convert the activation of conventional gene circuit-based sensors into a glucose output that can be read by off-the-shelf glucose meters. We show the development of new glucogenic reporter systems, multiplexed reporter outputs and detection of nucleic acid targets down to the low attomolar range. Using this glucose-meter interface, we demonstrate the detection of a small-molecule analyte; sample-to-result diagnostics for typhoid, paratyphoid A/B; and show the potential for pandemic response with nucleic acid sensors for SARS-CoV-2.

scholarly journals Demonstration of a Label-Free and Low-Cost Optical Cavity-Based Biosensor Using Streptavidin and C-Reactive Protein

Biosensors ◽  
2020 ◽  
Vol 11 (1) ◽  
pp. 4
Author(s):  
Donggee Rho ◽  
Seunghyun Kim
Keyword(s):  
Limit Of Detection ◽  
Point Of Care ◽  
Low Cost ◽  
Optical Cavity ◽  
Sample Volume ◽  
Small Sample ◽  
Label Free ◽  
C Reactive Protein ◽  
Reactive Protein ◽  
Cavity Structure

An optical cavity-based biosensor (OCB) has been developed for point-of-care (POC) applications. This label-free biosensor employs low-cost components and simple fabrication processes to lower the overall cost while achieving high sensitivity using a differential detection method. To experimentally demonstrate its limit of detection (LOD), we conducted biosensing experiments with streptavidin and C-reactive protein (CRP). The optical cavity structure was optimized further for better sensitivity and easier fluid control. We utilized the polymer swelling property to fine-tune the optical cavity width, which significantly improved the success rate to produce measurable samples. Four different concentrations of streptavidin were tested in triplicate, and the LOD of the OCB was determined to be 1.35 nM. The OCB also successfully detected three different concentrations of human CRP using biotinylated CRP antibody. The LOD for CRP detection was 377 pM. All measurements were done using a small sample volume of 15 µL within 30 min. By reducing the sensing area, improving the functionalization and passivation processes, and increasing the sample volume, the LOD of the OCB are estimated to be reduced further to the femto-molar range. Overall, the demonstrated capability of the OCB in the present work shows great potential to be used as a promising POC biosensor.

scholarly journals Pocket MUSE: an affordable, versatile and high-performance fluorescence microscope using a smartphone

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Yehe Liu ◽  
Andrew M. Rollins ◽  
Richard M. Levenson ◽  
Farzad Fereidouni ◽  
Michael W. Jenkins
Keyword(s):  
High Performance ◽  
Point Of Care ◽  
Low Cost ◽  
Consumer Electronics ◽  
Practical Implementation ◽  
The Novel ◽  
Point Of Care Diagnostics ◽  
Optical Module ◽  
Optical Configuration ◽  
User Friendly

AbstractSmartphone microscopes can be useful tools for a broad range of imaging applications. This manuscript demonstrates the first practical implementation of Microscopy with Ultraviolet Surface Excitation (MUSE) in a compact smartphone microscope called Pocket MUSE, resulting in a remarkably effective design. Fabricated with parts from consumer electronics that are readily available at low cost, the small optical module attaches directly over the rear lens in a smartphone. It enables high-quality multichannel fluorescence microscopy with submicron resolution over a 10× equivalent field of view. In addition to the novel optical configuration, Pocket MUSE is compatible with a series of simple, portable, and user-friendly sample preparation strategies that can be directly implemented for various microscopy applications for point-of-care diagnostics, at-home health monitoring, plant biology, STEM education, environmental studies, etc.

scholarly journals Nitrocellulose-bound achromopeptidase for point-of-care nucleic acid tests

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Georgios Chondrogiannis ◽  
Shirin Khaliliazar ◽  
Anna Toldrà ◽  
Pedro Réu ◽  
Mahiar M. Hamedi
Keyword(s):  
Nucleic Acid ◽  
Sample Preparation ◽  
Point Of Care ◽  
Dna Amplification ◽  
Recombinase Polymerase Amplification ◽  
Gram Positive ◽  
Gram Positive Bacteria ◽  
Enzymatic Function ◽  
Modern Biotechnology ◽  
Nucleic Acid Tests

AbstractEnzymes are the cornerstone of modern biotechnology. Achromopeptidase (ACP) is a well-known enzyme that hydrolyzes a number of proteins, notably proteins on the surface of Gram-positive bacteria. It is therefore used for sample preparation in nucleic acid tests. However, ACP inhibits DNA amplification which makes its integration difficult. Heat is commonly used to inactivate ACP, but it can be challenging to integrate heating into point-of-care devices. Here, we use recombinase polymerase amplification (RPA) together with ACP, and show that when ACP is immobilized on nitrocellulose paper, it retains its enzymatic function and can easily and rapidly be activated using agitation. The nitrocellulose-bound ACP does, however, not leak into the solution, preventing the need for deactivation through heat or by other means. Nitrocellulose-bound ACP thus opens new possibilities for paper-based Point-of-Care (POC) devices.

scholarly journals Slipdisc: a versatile sample preparation platform for point of care diagnostics

RSC Advances ◽  
2017 ◽  
Vol 7 (56) ◽  
pp. 35048-35054 ◽  
Author(s):  
I. Banerjee ◽  
T. Salih ◽  
H. Ramachandraiah ◽  
J. Erlandsson ◽  
T. Pettersson ◽  
...  
Keyword(s):  
Sample Preparation ◽  
Point Of Care ◽  
Preparation Technology ◽  
Minute Amount ◽  
Point Of Care Diagnostics

A novel POC sample preparation technology, “Slipdisc”, based rotational slipchip technology is presented. In operation, the hand-winded slipdisc platform uses a unique clockwork mechanism to manipulate minute amount of liquids.

Engineering Consideration for Emerging Essential Nucleic Acid Tests for Point-of-Care Diagnostics

2021 ◽  
Vol 4 ◽  
pp. 81-91
Author(s):  
Ethan P.M. LaRochelle ◽  
Amogha Tadimety
Keyword(s):  
Nucleic Acid ◽  
Point Of Care ◽  
Point Of Care Diagnostics ◽  
Nucleic Acid Tests

scholarly journals Nanomaterials for molecular signal amplification in electrochemical nucleic acid biosensing: recent advances and future prospects for point-of-care diagnostics

2020 ◽  
Vol 5 (1) ◽  
pp. 49-66 ◽  
Author(s):  
Léonard Bezinge ◽  
Akkapol Suea-Ngam ◽  
Andrew J. deMello ◽  
Chih-Jen Shih
Keyword(s):  
Nucleic Acid ◽  
Molecular Diagnostics ◽  
Signal Amplification ◽  
Point Of Care ◽  
Future Prospects ◽  
Electrochemical Biosensing ◽  
Recent Advances ◽  
Point Of Care Diagnostics ◽  
Molecular Signal

This account reviews the major amplification strategies utilizing nanomaterials in electrochemical biosensing for robust and sensitive molecular diagnostics.

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