scholarly journals All-in-One Dual CRISPR-Cas12a (AIOD-CRISPR) Assay: A Case for Rapid, Ultrasensitive and Visual Detection of Novel Coronavirus SARS-CoV-2 and HIV virus

2020 ◽  
Author(s):  
Xiong Ding ◽  
Kun Yin ◽  
Ziyue Li ◽  
Changchun Liu
Keyword(s):  
Disease Transmission ◽  
Visual Detection ◽  
Point Of Care ◽  
Detection Sensitivity ◽  
Transmission Risk ◽  
Assay Method ◽  
One Pot ◽  
Dna And Rna ◽  
Hiv Virus ◽  
Novel Coronavirus

AbstractA recent outbreak of novel coronavirus (SARS-CoV-2), the causative agent of COVID-19, has spread rapidly all over the world. Human immunodeficiency virus (HIV) is another deadly virus and causes acquired immunodeficiency syndrome (AIDS). Rapid and early detection of these viruses will facilitate early intervention and reduce disease transmission risk. Here, we present an All-In-One Dual CRISPR-Cas12a (termed “AIOD-CRISPR”) assay method for simple, rapid, ultrasensitive, one-pot, and visual detection of coronavirus SARS-CoV-2 and HIV virus. In our AIOD CRISPR assay, a pair of crRNAs was introduced to initiate dual CRISPR-Cas12a detection and improve detection sensitivity. The AIOD-CRISPR assay system was successfully utilized to detect nucleic acids (DNA and RNA) of SARS-CoV-2 and HIV with a sensitivity of few copies. Also, it was evaluated by detecting HIV-1 RNA extracted from human plasma samples, achieving a comparable sensitivity with real-time RT-PCR method. Thus, our method has a great potential for developing next-generation point-of-care molecular diagnostics.

scholarly journals All-in-One Dual CRISPR-Cas12a (AIOD-CRISPR) Assay: A Case for Rapid, Ultrasensitive and Visual Detection of Novel Coronavirus SARS-CoV-2 and HIV virus at the Point of Care

2020 ◽  
Author(s):  
Xiong Ding ◽  
Kun Yin ◽  
Ziyue Li ◽  
Rajesh V. Lalla ◽  
Enrique Ballesteros ◽  
...  
Keyword(s):  
Visual Detection ◽  
Point Of Care ◽  
Low Cost ◽  
Detection Sensitivity ◽  
Disease Spread ◽  
Assay Method ◽  
One Pot ◽  
Dna And Rna ◽  
Novel Coronavirus ◽  
Hiv 1

Abstract The recent outbreak of novel Coronavirus (SARS-CoV-2), the causative agent of COVID-19 disease, has spread rapidly all over the world. Human immunodeficiency virus (HIV) is another deadly virus and causes acquired immunodeficiency syndrome (AIDS). Rapid and early detection of these viruses will facilitate early intervention and prevent disease spread. Here, we present an All-In-One Dual CRISPR-Cas12a (termed "AIOD-CRISPR") assay method for simple, rapid, ultrasensitive, specific, one-pot, and visual detection of coronavirus SARS-CoV- 2 and HIV-1 virus. In our AIOD-CRISPR assay, a pair of crRNAs was introduced to initiate dual CRISPR-Cas12a-based detection and improve both detection sensitivity and fluorescence signals. The AIOD-CRISPR assay method was utilized to detect nucleic acids (DNA and RNA) of the SARS-CoV-2 and HIV-1 with a sensitivity of few copies. We validated our AIOD-CRISPR method by using COVID-19 swab samples and obtained consistent results with that of RT-PCR method. More importantly, we successfully demonstrated to use a low- cost hand warmer (~$ 0.3) as an incubator of our AIOD-CRISPR assay and detect COVID-19 patient samples within 20 minutes, enabling an instrument-free, visual detection of COVID-19 at the point of care. Thus, our method has significant potential for developing next-generation point-of-care molecular diagnostics.

scholarly journals Point of Care Diagnostic Devices for Rapid Detection of Novel Coronavirus (SARS-nCoV19) Pandemic: A Review

2021 ◽  
Vol 2 ◽  
Author(s):  
Vamkudoth Koteswara Rao
Keyword(s):  
Rapid Detection ◽  
Point Of Care ◽  
Transmission Risk ◽  
Causative Agents ◽  
Community Transmission ◽  
Severity Of The Disease ◽  
Novel Coronavirus ◽  
Approved Drugs ◽  
The Cost ◽  
Nucleic Acid Assays

Coronaviruses are recognized as causative agents of human diseases worldwide. In Wuhan, China, an outbreak of Severe acute respiratory syndrome novel Coronavirus (SARS-nCoV-2) was reported at the end of December 2019, causing 63 million COVID cases and 1.3 million deaths globally by 2 December, 2020. The transmission risk forecasts and the SARS-nCoV-2 epidemic pattern are progressive. Unfortunately, there is no specific FDA approved drugs or vaccines available currently to treat SARS-nCoV-2. In response to nCoV-2 spread, the rapid detection is crucial for estimating the severity of the disease and treatment of patients. Currently, there are several RT-PCR based diagnostic kits available for SARS-nCoV-2 detection, which are time-consuming, expensive, need advanced equipment facilities and trained personnel. The cost of diagnosis and the unavailability of sufficient test kits may prevent to check community transmission. Furthermore, expanding the testing facilities in asymptomatic cases in hotspots require more Point of Care (PoC) devices. Therefore, fast, inexpensive, and reliable methods of detection of SARS-nCoV-2 virus infection in humans is urgently required. The rapid and easy-to-use devices will facilitate onsite testing. In this review, nucleic acid assays, serological assays, multiplex assays, and PoC devices are discussed to understand various diagnostic approaches to reduce the spread and mortality rate in the future. Aptamer based detection is most specific, inexpensive and rapid detection of SARS-nCoV-2 without laboratory tools. To the best of our knowledge more than 900 SARS-nCoV-2 test kits are in pipeline, among 395 test kits are molecular bested test kits and only few test kits are developed using Aptamer technology https://www.finddx.org/covid-19/pipeline/.

scholarly journals Point-of-care testing for COVID-19 using SHERLOCK diagnostics

2020 ◽  
Author(s):  
Julia Joung ◽  
Alim Ladha ◽  
Makoto Saito ◽  
Michael Segel ◽  
Robert Bruneau ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Point Of Care ◽  
The Novel ◽  
One Pot ◽  
Viral Spread ◽  
Fluid Handling ◽  
Recent Outbreak ◽  
Alternative Approaches ◽  
Novel Coronavirus

AbstractThe recent outbreak of the novel coronavirus SARS-CoV-2, which causes COVID-19, can be diagnosed using RT-qPCR, but inadequate access to reagents and equipment has slowed disease detection and impeded efforts to mitigate viral spread. Alternative approaches based on combinations of isothermal amplification and CRISPR-mediated detection, such as the SHERLOCK (Specific High Sensitivity Enzymatic Reporter UnLOCKing) technique, offer reduced dependence on RT-qPCR equipment, but previously reported methods required multiple fluid handling steps, complicating their deployment outside clinical labs. Here we developed a simple test chemistry called STOP (SHERLOCK Testing in One Pot) for detecting SARS-CoV-2 in one hour that is suitable for point-of-care use. This simplified test, STOPCovid, provides sensitivity comparable to RT-qPCR-based SARS-CoV-2 tests and has a limit of detection of 100 copies of viral genome input in saliva or nasopharyngeal swabs per reaction. Using lateral flow readout, the test returns result in 70 minutes, and using fluorescence readout, the test returns result in 40 minutes. Moreover, we validated STOPCovid using nasopharyngeal swabs from COVID-19 patients and were able to correctly diagnose 12 positive and 5 negative patients out of 3 replicates. We envision that implementation of STOPCovid will significantly aid “test-trace-isolate” efforts, especially in low-resource settings, which will be critical for long-term public health safety and effective reopening of the society.

scholarly journals Ultrasensitive and visual detection of SARS-CoV-2 using all-in-one dual CRISPR-Cas12a assay

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Xiong Ding ◽  
Kun Yin ◽  
Ziyue Li ◽  
Rajesh V. Lalla ◽  
Enrique Ballesteros ◽  
...  
Keyword(s):  
Point Of Care ◽  
Low Cost ◽  
Disease Spread ◽  
Clinical Samples ◽  
Pcr Assay ◽  
One Pot ◽  
Nucleoprotein Gene ◽  
Protospacer Adjacent Motif ◽  
Recent Outbreak ◽  
Novel Coronavirus

Abstract The recent outbreak of novel coronavirus (SARS-CoV-2) causing COVID-19 disease spreads rapidly in the world. Rapid and early detection of SARS-CoV-2 facilitates early intervention and prevents the disease spread. Here, we present an All-In-One Dual CRISPR-Cas12a (AIOD-CRISPR) assay for one-pot, ultrasensitive, and visual SARS-CoV-2 detection. By targeting SARS-CoV-2’s nucleoprotein gene, two CRISPR RNAs without protospacer adjacent motif (PAM) site limitation are introduced to develop the AIOD-CRISPR assay and detect the nucleic acids with a sensitivity of few copies. We validate the assay by using COVID-19 clinical swab samples and obtain consistent results with RT-PCR assay. Furthermore, a low-cost hand warmer (~$0.3) is used as an incubator of the AIOD-CRISPR assay to detect clinical samples within 20 min, enabling an instrument-free, visual SARS-CoV-2 detection at the point of care. Thus, our method has the significant potential to provide a rapid, sensitive, one-pot point-of-care assay for SARS-CoV-2.

scholarly journals All-in-one dual CRISPR-Cas12a (AIOD-CRISPR) assay protocol for SARS-CoV-2 detection

2020 ◽  
Author(s):  
Xiong Ding ◽  
Kun Yin ◽  
Ziyue Li ◽  
Rajesh V. Lalla ◽  
Maroun M. Sfeir ◽  
...  
Keyword(s):  
Sample Preparation ◽  
Fluorescence Detection ◽  
Visual Detection ◽  
Imaging System ◽  
Point Of Care ◽  
Acid Extraction ◽  
Nucleic Acid Extraction ◽  
One Pot ◽  
Reaction Systems ◽  
Assay Protocol

Abstract This protocol presents the all-in-one dual CRISPR-Cas12a (AIOD-CRISPR) assay to ultra-sensitively and visually detect SARS-CoV-2. The procedure of AIOD-CRISPR assay typically consists of three parts including sample preparation, AIOD-CRISPR reaction, and fluorescence detection. Sample preparation involves the synthetic RNA preparation and the nucleic acid extraction from SARS-CoV-2 samples. The prepared nucleic acids were then added into the AIOD-CRISPR reaction systems as templates, followed by incubation at 37°C for 20-40 min. After incubation, visual detection was immediately conducted by placing the tubes in a portable LED blue transilluminator (Maestrogen UltraSlim) or the ChemiDoc™ MP Imaging System (Bio-Rad) with its built-in UV channel. In addition to endpoint visual detection, real-time fluorescence detection was also available for AIOD-CRISPR assay. This protocol is helpful for applying AIOD-CRISPR assay to rapid, sensitive, one-pot point-of-care SARS-CoV-2 detection.

Ferromagnetic Resonance Biosensor for Homogeneous and Volumetric Detection of DNA

2017 ◽  
Author(s):  
Bo Tian ◽  
Peter Svedlindh ◽  
Mattias Strömberg ◽  
Erik Wetterskog
Keyword(s):  
Magnetic Nanoparticles ◽  
Ferromagnetic Resonance ◽  
Limit Of Detection ◽  
Point Of Care ◽  
Rolling Circle Amplification ◽  
Resonance Field ◽  
Isothermal Amplification ◽  
Detection Sensitivity ◽  
Serum Samples ◽  
Rolling Circle

In this work, we demonstrate for the first time, a ferromagnetic resonance (FMR) based homogeneous and volumetric biosensor for magnetic label detection. Two different isothermal amplification methods, <i>i.e.</i>, rolling circle amplification (RCA) and loop-mediated isothermal amplification (LAMP) are adopted and combined with a standard electron paramagnetic resonance (EPR) spectrometer for FMR biosensing. For RCA-based FMR biosensor, binding of RCA products of a synthetic Vibrio cholerae target DNA sequence gives rise to the formation of aggregates of magnetic nanoparticles. Immobilization of nanoparticles within the aggregates leads to a decrease of the net anisotropy of the system and a concomitant increase of the resonance field. A limit of detection of 1 pM is obtained with an average coefficient of variation of 0.16%, which is superior to the performance of other reported RCA-based magnetic biosensors. For LAMP-based sensing, a synthetic Zika virus target oligonucleotide is amplified and detected in 20% serum samples. Immobilization of magnetic nanoparticles is induced by their co-precipitation with Mg<sub>2</sub>P<sub>2</sub>O<sub>7</sub> (a by-product of LAMP) and provides a detection sensitivity of 100 aM. The fast measurement, high sensitivity and miniaturization potential of the proposed FMR biosensing technology makes it a promising candidate for designing future point-of-care devices.<br>

scholarly journals ASE Statement on Point-of-Care Ultrasound during the 2019 Novel Coronavirus Pandemic

2020 ◽  
Vol 33 (6) ◽  
pp. 670-673 ◽  
Author(s):  
Amer M. Johri ◽  
Benjamin Galen ◽  
James N. Kirkpatrick ◽  
Michael Lanspa ◽  
Sharon Mulvagh ◽  
...  
Keyword(s):  
Point Of Care ◽  
Point Of Care Ultrasound ◽  
Novel Coronavirus

The performance of a new point-of-care HIV virus load technology to identify patients failing antiretroviral treatment

2020 ◽  
Vol 122 ◽  
pp. 104212
Author(s):  
Diana Mariani ◽  
Marcelo C.V.M. de Azevedo ◽  
Isabelle Vasconcellos ◽  
Luiz Ribeiro ◽  
Cassia Alves ◽  
...  
Keyword(s):  
Antiretroviral Treatment ◽  
Point Of Care ◽  
Virus Load ◽  
Hiv Virus

scholarly journals Livestock movement informs the risk of disease spread in traditional production systems in East Africa

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Divine Ekwem ◽  
Thomas A. Morrison ◽  
Richard Reeve ◽  
Jessica Enright ◽  
Joram Buza ◽  
...  
Keyword(s):  
Disease Transmission ◽  
Production Systems ◽  
Control Strategies ◽  
Herd Size ◽  
Transmission Risk ◽  
Disease Spread ◽  
Transmission Potential ◽  
Spatial And Temporal Scales ◽  
Transmission Distance ◽  
Risk Of Disease

AbstractIn Africa, livestock are important to local and national economies, but their productivity is constrained by infectious diseases. Comprehensive information on livestock movements and contacts is required to devise appropriate disease control strategies; yet, understanding contact risk in systems where herds mix extensively, and where different pathogens can be transmitted at different spatial and temporal scales, remains a major challenge. We deployed Global Positioning System collars on cattle in 52 herds in a traditional agropastoral system in western Serengeti, Tanzania, to understand fine-scale movements and between-herd contacts, and to identify locations of greatest interaction between herds. We examined contact across spatiotemporal scales relevant to different disease transmission scenarios. Daily cattle movements increased with herd size and rainfall. Generally, contact between herds was greatest away from households, during periods with low rainfall and in locations close to dipping points. We demonstrate how movements and contacts affect the risk of disease spread. For example, transmission risk is relatively sensitive to the survival time of different pathogens in the environment, and less sensitive to transmission distance, at least over the range of the spatiotemporal definitions of contacts that we explored. We identify times and locations of greatest disease transmission potential and that could be targeted through tailored control strategies.

scholarly journals Addressing the COVID-19 transmission in inner Brazil by a mathematical model

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
G. B. Almeida ◽  
T. N. Vilches ◽  
C. P. Ferreira ◽  
C. M. C. B. Fortaleza
Keyword(s):  
Mathematical Model ◽  
Disease Transmission ◽  
Abiotic Factors ◽  
Severe Pneumonia ◽  
Mitigation Strategies ◽  
Social Distancing ◽  
Control Disease ◽  
Set Up ◽  
Novel Coronavirus ◽  
Public Health Strategies

AbstractIn 2020, the world experienced its very first pandemic of the globalized era. A novel coronavirus, SARS-CoV-2, is the causative agent of severe pneumonia and has rapidly spread through many nations, crashing health systems and leading a large number of people to death. In Brazil, the emergence of local epidemics in major metropolitan areas has always been a concern. In a vast and heterogeneous country, with regional disparities and climate diversity, several factors can modulate the dynamics of COVID-19. What should be the scenario for inner Brazil, and what can we do to control infection transmission in each of these locations? Here, a mathematical model is proposed to simulate disease transmission among individuals in several scenarios, differing by abiotic factors, social-economic factors, and effectiveness of mitigation strategies. The disease control relies on keeping all individuals’ social distancing and detecting, followed by isolating, infected ones. The model reinforces social distancing as the most efficient method to control disease transmission. Moreover, it also shows that improving the detection and isolation of infected individuals can loosen this mitigation strategy. Finally, the effectiveness of control may be different across the country, and understanding it can help set up public health strategies.

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