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

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.

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

Molecular Systems Design & Engineering ◽

10.1039/c9me00135b ◽

2020 ◽

Vol 5 (1) ◽

pp. 49-66 ◽

Cited By ~ 11

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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Rapid point‐of‐care diagnostics for the detection of Chlamydia pecorum in koalas ( Phascolarctos cinereus ) using loop‐mediated isothermal amplification without nucleic acid purification

MicrobiologyOpen ◽

10.1002/mbo3.916 ◽

2019 ◽

Vol 8 (12) ◽

Cited By ~ 1

Author(s):

Lyndal S. Hulse ◽

Sean McDonald ◽

Stephen D. Johnston ◽

Kenneth W. Beagley

Keyword(s):

Nucleic Acid ◽

Point Of Care ◽

Isothermal Amplification ◽

Phascolarctos Cinereus ◽

Loop Mediated Isothermal Amplification ◽

Chlamydia Pecorum ◽

Point Of Care Diagnostics ◽

Nucleic Acid Purification

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Nucleic acid aptamers: ideal reagents for point-of-care diagnostics?

Faraday Discussions ◽

10.1039/c005487a ◽

2011 ◽

Vol 149 ◽

pp. 49-61 ◽

Cited By ~ 15

Author(s):

Anthony E. G. Cass ◽

Yangyang Zhang

Keyword(s):

Nucleic Acid ◽

Point Of Care ◽

Nucleic Acid Aptamers ◽

Point Of Care Diagnostics

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A simple check valve for microfluidic point of care diagnostics

Lab on a Chip ◽

10.1039/c6lc01104g ◽

2016 ◽

Vol 16 (22) ◽

pp. 4436-4444 ◽

Cited By ~ 9

Author(s):

C. S. Ball ◽

R. F. Renzi ◽

A. Priye ◽

R. J. Meagher

Keyword(s):

Nucleic Acid ◽

Point Of Care ◽

Check Valve ◽

Nucleic Acid Amplification ◽

Nucleic Acid Amplification Testing ◽

Point Of Care Diagnostics

Laser cut microfluidic check valves enable staged reagent delivery, pumping, and point of care nucleic acid amplification testing.

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Current State of Commercial Point-of-Care Nucleic Acid Tests for Infectious Diseases

The Analyst ◽

10.1039/d0an01988g ◽

2021 ◽

Author(s):

Jane Zhang ◽

Andrew T. Bender ◽

David S. Boyle ◽

Paul K Drain ◽

Jonathan D. Posner

Keyword(s):

Nucleic Acid ◽

Infectious Diseases ◽

Point Of Care ◽

Current State ◽

Nucleic Acid Tests

The COVID-19 pandemic has put the spotlight on the urgent need for integrated nucleic acid tests (NATs) for infectious diseases, especially those that can be used near patient (“point-of-care”, POC),...

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

Scientific Reports ◽

10.1038/s41598-019-52922-y ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 4

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.

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Current Nucleic Acid Extraction Methods and Their Implications to Point-of-Care Diagnostics

BioMed Research International ◽

10.1155/2017/9306564 ◽

2017 ◽

Vol 2017 ◽

pp. 1-13 ◽

Cited By ~ 56

Author(s):

Nasir Ali ◽

Rita de Cássia Pontello Rampazzo ◽

Alexandre Dias Tavares Costa ◽

Marco Aurelio Krieger

Keyword(s):

Nucleic Acid ◽

Extraction Method ◽

Point Of Care ◽

Extraction Methods ◽

Vital Role ◽

Acid Extraction ◽

Nucleic Acid Extraction ◽

Advantages And Disadvantages ◽

Point Of Care Diagnostics ◽

Working Principle

Nucleic acid extraction (NAE) plays a vital role in molecular biology as the primary step for many downstream applications. Many modifications have been introduced to the original 1869 method. Modern processes are categorized into chemical or mechanical, each with peculiarities that influence their use, especially in point-of-care diagnostics (POC-Dx). POC-Dx is a new approach aiming to replace sophisticated analytical machinery with microanalytical systems, able to be used near the patient, at thepoint of careorpoint of need. Although notable efforts have been made, a simple and effective extraction method is still a major challenge for widespread use of POC-Dx. In this review, we dissected the working principle of each of the most common NAE methods, overviewing their advantages and disadvantages, as well their potential for integration in POC-Dx systems. At present, it seems difficult, if not impossible, to establish a procedure which can be universally applied to POC-Dx. We also discuss the effects of the NAE chemicals upon the main plastic polymers used to mass produce POC-Dx systems. We end our review discussing the limitations and challenges that should guide the quest for an efficient extraction method that can be integrated in a POC-Dx system.

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CRISPR-Based COVID-19 Testing: Toward Next-Generation Point-of-Care Diagnostics

Frontiers in Cellular and Infection Microbiology ◽

10.3389/fcimb.2021.663949 ◽

2021 ◽

Vol 11 ◽

Author(s):

Uyanga Ganbaatar ◽

Changchun Liu

Keyword(s):

Nucleic Acid ◽

Point Of Care ◽

High Sensitivity ◽

Detection Methods ◽

Molecular Testing ◽

Practical Applications ◽

Food Industries ◽

Resource Limited ◽

Point Of Care Diagnostics ◽

Agriculture And Food

As the COVID-19 pandemic continues, people are becoming infected at an alarming rate, individuals are unknowingly spreading disease, and more lives are lost every day. There is an immediate need for a simple, rapid, early and sensitive point-of-care testing for COVID-19 disease. However, current testing approaches do not meet such need. Recently, clustered regularly interspaced short palindromic repeats (CRISPR)-based detection methods have received substantial attention for nucleic acid-based molecular testing due to their simplicity, high sensitivity and specificity. This review explores the various CRISPR-based COVID-19 detection methods and related diagnostic devices. As with any emerging technology, CRISPR/Cas-based nucleic acid testing methods have several challenges that must be overcome for practical applications in clinics and hospitals. More importantly, these detection methods are not limited to COVID-19 but can be applied to detect any type of pathogen, virus, and fungi that may threaten humans, agriculture, and food industries in resource-limited settings. CRISPR/Cas-based detection methods have the potential to become simpler, more reliable, more affordable, and faster in the near future, which is highly important for achieving point-of-care diagnostics.

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