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1.
ACS Sens ; 6(11): 3957-3966, 2021 11 26.
Article in English | MEDLINE | ID: covidwho-1493024

ABSTRACT

The development of an extensive toolkit for potential point-of-care diagnostics that is expeditiously adaptable to new emerging pathogens is of critical public health importance. Recently, a number of novel CRISPR-based diagnostics have been developed to detect SARS-CoV-2. Herein, we outline the development of an alternative CRISPR nucleic acid diagnostic utilizing a Cas13d ribonuclease derived from Ruminococcus flavefaciens XPD3002 (CasRx) to detect SARS-CoV-2, an approach we term SENSR (sensitive enzymatic nucleic acid sequence reporter) that can detect attomolar concentrations of SARS-CoV-2. We demonstrate 100% sensitivity in patient-derived samples by lateral flow and fluorescence readout with a detection limit of 45 copy/µL. This technology expands the available nucleic acid diagnostic toolkit, which can be adapted to combat future pandemics.

2.
Front Neurol ; 12: 743110, 2021.
Article in English | MEDLINE | ID: covidwho-1485083

ABSTRACT

Objective: We conducted a survey to assess vaccination coverage, vaccination willingness, and variables associated with vaccination hesitancy to provide evidence on coronavirus disease (COVID-19) vaccination strategies. Methods: This anonymous questionnaire study conducted a multicenter, cross-sectional survey of outpatients and inpatients with epilepsy (PWE) registered in epilepsy clinics, in 2021, in 10 hospitals in seven cities of Shandong Province. Results: A total of 600 questionnaires were distributed, and 557 valid questionnaires were returned. A total of 130 people were vaccinated against COVID-19. Among 427 unvaccinated participants, 69.32% (296/427) were willing to receive the COVID-19 vaccine in the future, and the remaining 30.68% (131/427) were unwilling to receive vaccination. Most (89.9%) of the participants believed that the role of vaccination was crucial in response to the spread of COVID-19. A significant association was found between willingness to receive the COVID-19 vaccine and the following variables: age, marital status, level of education, occupation, residence, seizure type, and seizure control after antiepileptic drug therapy. It is noteworthy that education level, living in urban areas, and seizure freedom were significantly related to willingness to receive COVID-19 vaccination. Conclusions: Vaccination is a key measure for the prevention and control of COVID-19, and most PWE are willing to be vaccinated. Vaccine safety, effectiveness, and accessibility are essential in combatting vaccine hesitation and increasing vaccination rates.

4.
Int J Immunopathol Pharmacol ; 35: 20587384211048567, 2021.
Article in English | MEDLINE | ID: covidwho-1463208

ABSTRACT

BACKGROUND: Coronavirus disease 2019 (COVID-19) had become a worldwide health threat. Early prediction of the severity of COVID-19 patients was important for reducing death rate and controlling this disease. METHODS AND MATERIALS: A total of 301 patients confirmed with COVID-19 in Wuhan from 8 February to 10 April 2020 were included. Clinical data were collected and analyzed. Diagnostic and prognostic utility of blood cell counts and lymphocyte subsets in COVID-19 patients were investigated. The receiver operator characteristic curve (ROC) was used in discriminating the mild and severe/critical cases. RESULTS: There were difference in blood cell counts and lymphocyte subsets among mild, severe and critical patients, which were also influenced by comorbidities and duration of disease. The area under the ROC of lymphocyte, CD3+ T cells, CD4+ T cells, and CD8+ T cells were 0.718, 0.721, 0.718, and 0.670, which were higher than that of other hematological parameters. The optimal threshold was 1205, 691, 402, and 177 per µl, respectively. Patients with higher counts of lymphocyte, CD3+ T cells, CD4+ T cells, or CD8+ T cells were correlated with shorter length of stay in hospital (p < 0.05). Multivariable Cox regression analysis showed disease severity, CD3+ T cells counts and time when the nucleic acid turned negative were independent risk factors for in-hospital death of COVID-19 patients (p < 0.05). CONCLUSION: Blood cell counts and lymphocyte subsets correlated with severity of COVID-19.


Subject(s)
COVID-19/immunology , Lymphocyte Subsets/immunology , Adult , Aged , Aged, 80 and over , CD4 Lymphocyte Count , COVID-19/diagnosis , COVID-19/mortality , COVID-19/virology , China , Female , Hospital Mortality , Host-Pathogen Interactions , Humans , Lymphocyte Subsets/virology , Male , Middle Aged , Phenotype , Predictive Value of Tests , Prognosis , Retrospective Studies , SARS-CoV-2/immunology , SARS-CoV-2/pathogenicity , Severity of Illness Index , Time Factors , Young Adult
5.
Clin Infect Dis ; 2020 Nov 09.
Article in English | MEDLINE | ID: covidwho-1387786

ABSTRACT

BACKGROUND: We evaluated an inactivated SARS-CoV-2 vaccine for immunogenicity and safety in adults aged 18-59 years. METHODS: In this randomized, double-blinded and controlled trial, healthy adults received a medium (MD) or a high dose (HD) of the vaccine at an interval of either 14 days or 28 days. Neutralizing antibody (NAb) and anti-S and anti-N antibodies were detected at different times, and adverse reactions were monitored for 28 days after full immunization. RESULTS: A total of 742 adults were enrolled in the immunogenicity and safety analysis. Among subjects in the 0, 14 procedure, the seroconversion rates of NAb in MD and HD groups were 89% and 96% with GMTs of 23 and 30, respectively, at day 14 and 92% and 96% with GMTs of 19 and 21, respectively at day 28 after immunization. Anti-S antibodies had GMTs of 1883 and 2370 in MD and 2295 and 2432 in HD group. Anti-N antibodies had GMTs of 387 and 434 in MD group and 342 and 380 in HD group. Among subjects in the 0, 28 procedure, seroconversion rates for NAb at both doses were both 95% with GMTs of 19 at day 28 after immunization. Anti-S antibodies had GMTs of 937 and 929 for MD and HD group, and anti-N antibodies had GMTs of 570 and 494 for MD and HD group, respectively. No serious adverse events were observed during the study period. CONCLUSION: Adults vaccinated with inactivated SARS-CoV-2 vaccine had NAb as well as anti-S/N antibody, and had a low rate of adverse reactions. CLINICAL TRIALS REGISTRATION: NCT04412538.

6.
Chinese Journal of Emergency Medicine ; 29(5):694-699, 2020.
Article in Chinese | CAB Abstracts | ID: covidwho-1328149

ABSTRACT

Objective: To explore the efficacy and case selection of non-invasive positive pressure ventilation (NIPPV) in the treatment of acute respiratory distress syndrome (ARDS) caused by viral pneumonia.

7.
Clin Respir J ; 15(11): 1158-1167, 2021 Nov.
Article in English | MEDLINE | ID: covidwho-1309757

ABSTRACT

INTRODUCTION: The novel coronavirus disease 2019 (COVID-19) could cause physical and psychological dysfunction in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infected patients. Pulmonary telerehabilitation is strongly recommended due to the contagious nature of the disease, but its efficacy and benefit need to be comprehensively evaluated. METHODS: An 8-week pulmonary telerehabilitation programme is designed in a real-world and prospective manner, aiming to assess the efficacy of remote rehabilitation among discharged patients with COVID-19 from three designated hospitals in the epicenter of China (Wuhan City). The telerehabilitation programme includes physicians and physiotherapists remotely guided training (online) for 2 weeks, and patient self-managed rehabilitation (offline) for 6 weeks with assistance of information technology and digital device. The main rehabilitation intervention includes breathing exercise, respiratory muscle training, aerobic exercise, and resistance training. The primary outcome is 6-min walk distance, and secondly pulmonary function, respiratory muscle strength, physical activity, symptom, psychological status and quality of life, etc. will also be assessed. Patients will be followed up periodically at week two, four, eight after enrolment and at month one, three, six and 12 after the rehabilitation accomplished, respectively. Current study has been approved by the ethics committee of China-Japan Friendship Hospital and three participating centers in Wuhan City. DISCUSSION: We anticipate that the health status of discharged patients with COVID-19 could be improved through this systemic pulmonary telerehabilitation programme. The interdisciplinary remote mode of rehabilitation could be an optimal way in management of emerging respiratory tract infectious disease. TRIAL REGISTRATION: ClinicalTrials.gov, NCT04368793.


Subject(s)
COVID-19 , Telerehabilitation , Humans , Patient Discharge , Prospective Studies , Quality of Life , SARS-CoV-2
8.
Eur J Med Res ; 26(1): 61, 2021 Jun 24.
Article in English | MEDLINE | ID: covidwho-1282268

ABSTRACT

BACKGROUND: The COVID-19 pandemic occurred and rapidly spread around the world. Some online dashboards have included essential features on a world map. However, only transforming data into visualizations for countries/regions is insufficient for the public need. This study aims to (1) develop an algorithm for classifying countries/regions into four quadrants inn GSM and (2) design an app for a better understanding of the COVID-19 situation. METHODS: We downloaded COVID-19 outbreak numbers daily from the Github website, including 189 countries/regions. A four-quadrant diagram was applied to present the classification of each country/region using Google Maps run on dashboards. A novel presentation scheme was used to identify the most struck entities by observing (1) the multiply infection rate (MIR) and (2) the growth trend in the recent 7 days. Four clusters of the COVID-19 outbreak were dynamically classified. An app based on a dashboard aimed at public understanding of the outbreak types and visualizing of the COVID-19 pandemic with Google Maps run on dashboards. The absolute advantage coefficient (AAC) was used to measure the damage hit by COVID-19 referred to the next two countries severely hit by COVID-19. RESULTS: We found that the two hypotheses were supported: India (i) is in the increasing status as of April 28, 2021; (ii) has a substantially higher ACC(= 0.81 > 0.70), and (iii) has a substantially higher ACC(= 0.66 < 0.70) as of May 17, 2021. CONCLUSION: Four clusters of the COVID-19 outbreak were dynamically classified online on an app making the public understand the outbreak types of COVID-19 pandemic shown on dashboards. The app with GSM and AAC is recommended for researchers in other disease outbreaks, not just limited to COVID-19.


Subject(s)
Algorithms , COVID-19/epidemiology , COVID-19/transmission , Global Health/statistics & numerical data , Models, Statistical , SARS-CoV-2/isolation & purification , Humans , India/epidemiology
10.
Biochem Biophys Res Commun ; 529(2): 251-256, 2020 08 20.
Article in English | MEDLINE | ID: covidwho-1220683

ABSTRACT

The nucleocapsid protein is significant in the formation of viral RNA of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), accounting for the largest proportion of viral structural proteins. Here, we report for the first time that the 11S proteasomal activator PA28γ regulates the intracellular abundance of the SARS-CoV-2 N protein (nCoV N). Furthermore, we have identified proteasome activator PA28γ as a nCoV N binding protein by co-immunoprecipitation assay. As a result of their interaction, nCoV N could be degraded by PA28γ-20S in vitro degradation assay. This was also demonstrated by blocking de novo protein synthesis with cycloheximide. The stability of nCoV N in PA28γ-knockout cells was greater than in PA28γ-wildtype cells. Notably, immunofluorescence staining revealed that knockout of the PA28γ gene in cells led to the transport of nCoV N from the nucleus to the cytoplasm. Overexpression of PA28γ enhanced proteolysis of nCoV N compared to that in PA28γ-N151Y cells containing a dominant-negative PA28γ mutation, which reduced this process. These results suggest that PA28γ binding is important in regulating 20S proteasome activity, which in turn regulates levels of the critical nCoV N nucleocapsid protein of SARS-CoV-2, furthering our understanding of the pathogenesis of COVID-19.


Subject(s)
Autoantigens/metabolism , Betacoronavirus/metabolism , Coronavirus Infections/metabolism , Nucleocapsid Proteins/metabolism , Pneumonia, Viral/metabolism , Proteasome Endopeptidase Complex/metabolism , Proteolysis , COVID-19 , Coronavirus Infections/virology , Coronavirus Nucleocapsid Proteins , HEK293 Cells , Humans , In Vitro Techniques , Pandemics , Phosphoproteins , Pneumonia, Viral/virology , Protein Binding , Protein Stability , Protein Transport , SARS-CoV-2
11.
J Med Virol ; 93(3): 1732-1738, 2021 03.
Article in English | MEDLINE | ID: covidwho-1196496

ABSTRACT

The coronavirus disease 2019 pandemic caused by severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) had led to a serious public health crisis, and no specific treatments or vaccines are available yet. A nucleocapsid protein (NP)-based enzyme-linked immunosorbent assay (ELISA) detection method is not only important in disease diagnosis, but is required for the evaluation of vaccine efficacy during the development of an inactivated SARS-CoV-2 vaccine. In this study, we expressed both the NP and N-terminally truncated NP (ΔN-NP) of SARS-CoV-2 in an Escherichia coli expression system and described the purification of the soluble recombinant NP and ΔN-NP in details. The identities of the NP and ΔN-NP were confirmed with mass spectrometry. We then used immunoglobulin G detection ELISAs to compare the sensitivity of NP and ΔN-NP in detecting anti-SARS-CoV-2 antibodies. ΔN-NP showed greater sensitivity than NP in the analysis of serially diluted sera from mice and rabbits vaccinated with inactive SARS-CoV-2 and in human sera diluted 1:400. ΔN-NP showed a positive detection rate similar to that of the SARS-CoV-2 S protein in human sera. We conclude that ΔN-NP is a better serological marker than NP for evaluating the immunogenicity of inactivated SARS-CoV-2.


Subject(s)
Antibodies, Viral/immunology , COVID-19 Vaccines/immunology , Coronavirus Nucleocapsid Proteins/immunology , SARS-CoV-2/immunology , Spike Glycoprotein, Coronavirus/immunology , Vaccines, Inactivated/immunology , Animals , COVID-19/prevention & control , Coronavirus Nucleocapsid Proteins/genetics , Humans , Mice , Mice, Inbred BALB C , Phosphoproteins/genetics , Phosphoproteins/immunology , Rabbits , SARS-CoV-2/genetics , Sequence Deletion/genetics , Sequence Deletion/immunology , Spike Glycoprotein, Coronavirus/genetics
12.
Technol Soc ; 64: 101514, 2021 Feb.
Article in English | MEDLINE | ID: covidwho-1185285

ABSTRACT

The continuous spread of the COVID-19 pandemic is causing people to feel anxiety and stress. This study constructs a four-layer research model to examine how a 360° virtual tour can reduce people's psychological stress through two types of presence (the sense of presence and telepresence) and affective-motivational states (enjoyment and involvement) in this extraordinary period of the COVID-19 pandemic. In order to test the moderating effect of involvement, partial least squares (PLS) analysis is employed to analyse valid data collected from 235 individuals. The results of this study indicate that telepresence has a higher impact in generating affective-motivational states than the sense of presence. Among the factors, enjoyment shows the highest effect on satisfaction with the 360° virtual tour experience and stress reduction; involvement moderates the effect of telepresence on satisfaction with the 360° virtual tour experience. This study also contributes to virtual reality research by distinguishing the concepts of 'sense of presence' and 'telepresence' as well as demonstrating the mechanisms whereby virtual reality technology influences people's psychological well-being. Timely recommendations are provided for people in order to reduce psychological stress during and after COVID-19 pandemic.

13.
The International Journal of Electrical Engineering & Education ; : 00207209211003270, 2021.
Article in English | Sage | ID: covidwho-1153835

ABSTRACT

Under the circumstance of COVID-19, online education has become an inevitable choice and a mainstream form of education in various countries. Intelligent Tutoring Systems ( ITSs ) are a critical category of a carrier of online education which refers to using Artificial Intelligence Technology ( AIT ) to realize distance teaching and learning through integrating multiple resources, assist teachers to implement online classes and help students learn courses online, etc. The target of this study is to examine the determinants that explain college students? adoption of ITSs based on TRA and TAM under the era of COVID-19 in China.. This study targets college students who have used ITSs over the previous year to learn courses. In total, 590 valid samples were collected. Partial least squares ( PLS ) approach is employed to inspect the determinants of college students? acceptance of using ITSs. The results of the analysis indicate that perceived ease of use, perceived usefulness, subjective norm, college students? attitude and political influence significantly affect college students? behavioral intention towards using ITSs. In addition, perceived usefulness, perceived ease of use, and political influence also affect college students? attitude toward using ITSs. And consisitent with previous research, perceived ease of use has a significant effects on perceived usefulness. This study enriches the TRA and TAM theories by incorporating a new variable political influence for researchers to consider in the future. In addition, it makes several suggestions for system developers to improve system functions regarding its ease of use and for content manufactures to pay attention to quality regarding usefulness. Recommendations are provided to colleges, universities and education authorities to rethink their roles in reforming online education through ITSs.

14.
Cell Rep ; 34(4): 108666, 2021 01 26.
Article in English | MEDLINE | ID: covidwho-1064915

ABSTRACT

Although vaccines against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are under development, the antigen epitopes on the virus and their immunogenicity are poorly understood. Here, we simulate the 3D structures and predict the B cell epitopes on the spike (S), envelope (E), membrane (M), and nucleocapsid (N) proteins of SARS-CoV-2 using structure-based approaches and validate epitope immunogenicity by immunizing mice. Almost all 33 predicted epitopes effectively induce antibody production, six of these are immunodominant epitopes in individuals, and 23 are conserved within SARS-CoV-2, SARS-CoV, and bat coronavirus RaTG13. We find that the immunodominant epitopes of individuals with domestic (China) SARS-CoV-2 are different from those of individuals with imported (Europe) SARS-CoV-2, which may be caused by mutations on the S (G614D) and N proteins. Importantly, we find several epitopes on the S protein that elicit neutralizing antibodies against D614 and G614 SARS-CoV-2, which can contribute to vaccine design against coronaviruses.


Subject(s)
Coronavirus Nucleocapsid Proteins/immunology , Epitopes, B-Lymphocyte/immunology , SARS-CoV-2/immunology , Spike Glycoprotein, Coronavirus/immunology , Viral Matrix Proteins/immunology , Viroporin Proteins/immunology , Adolescent , Adult , Aged , Animals , Antibodies, Neutralizing/immunology , Antibodies, Viral/immunology , Antigens, Viral/immunology , COVID-19/immunology , COVID-19/therapy , COVID-19 Vaccines/immunology , Child , Epitopes, B-Lymphocyte/metabolism , Female , Humans , Male , Mice , Mice, Inbred BALB C , Middle Aged , Young Adult
15.
The International Journal of Electrical Engineering & Education ; : 0020720920984001, 2021.
Article in English | Sage | ID: covidwho-1039961

ABSTRACT

During the COVID-19 pandemic, Online Tutoring Platforms (OTPs) have been applied extensively in education in China. The aim of this study is to investigate the determinants that influence college students? behavioural intention of using OTPs to acquire knowledge. This study also explores how students? attitude change in using OTPs five months after the outbreak of the COVID-19 pandemic based on a hierarchical component model which is combined by the Social Impact Theory (SIT) and Unified Theory of Acceptance and Use of Technology (UTAUT). Partial least squares (PLS) analysis is employed to analyse the data collected from 1133 students in Mainland China. The results of the analysis indicate that social impact consists of three dimensions (compliance, identification, internalization) significantly influences college students? attitude toward OTPs and further affects college students? behavioural intention toward OTPs usage. Furthermore, performance expectancy and effort expectancy also positively affect students? behavioural intention toward using OTPs to acquire knowledge. This study makes several suggestions for universities to encourage students using OTPs to cope with the situation of Covid-19 pandemic and for educators to promote online tutoring for reforming universities in the future.

16.
Shock ; 56(3): 360-367, 2021 09 01.
Article in English | MEDLINE | ID: covidwho-1028641

ABSTRACT

PURPOSE: Rhabdomyolysis (RM) has been associated with many viral infectious diseases, and associated with poor outcomes. We aim to evaluate the clinical features and outcomes of RM in patients with coronavirus disease 2019 (COVID-19). METHOD: This was a single-center, retrospective, cohort study of 1,014 consecutive hospitalized patients with confirmed COVID-19 at the Huoshenshan Hospital in Wuhan, China, between February 17 and April 12, 2020. RESULTS: The overall incidence of RM was 2.2%. Compared with patients without RM, those with RM tended to have a higher risk of deterioration. Patients with RM also constituted a greater percentage of patients admitted to the intensive care unit (90.9% vs. 5.3%, P < 0.001) and a greater percentage of patients undergoing mechanical ventilation (86.4% vs. 2.7% P < 0.001). Moreover, patients with RM had laboratory test abnormalities, including the presence of markers of inflammation, activation of coagulation, and kidney injury. Patients with RM also had a higher risk of in-hospital death (P < 0.001). Cox's proportional hazard regression model analysis confirmed that RM indicators, including peak creatine kinase levels > 1,000 IU/L (HR = 6.46, 95% CI: 3.02-13.86) and peak serum myoglobin concentrations > 1,000 ng/mL (HR = 9.85, 95% CI: 5.04-19.28), were independent risk factors for in-hospital death. Additionally, patients with COVID-19 that developed RM tended to have delayed viral clearance. CONCLUSION: RM might be an important contributing factor to adverse outcomes in COVID-19 patients. The early detection and effective intervention of RM may help reduce mortality among COVID-19 patients.


Subject(s)
COVID-19/complications , COVID-19/mortality , Hospital Mortality , Rhabdomyolysis/complications , Rhabdomyolysis/mortality , Adolescent , Adult , Aged , Aged, 80 and over , China/epidemiology , Female , Hospitalization , Humans , Incidence , Intensive Care Units , Male , Middle Aged , Muscle, Skeletal/physiopathology , Proportional Hazards Models , Respiration, Artificial , Retrospective Studies , SARS-CoV-2 , Treatment Outcome , Young Adult
17.
Small Methods ; 5(2): 2001031, 2021 Feb 15.
Article in English | MEDLINE | ID: covidwho-986422

ABSTRACT

The ongoing corona virus disease 2019 (COVID-19) pandemic, caused by SARS-CoV-2 infection, has resulted in hundreds of thousands of deaths. Cellular entry of SARS-CoV-2, which is mediated by the viral spike protein and ACE2 receptor, is an essential target for the development of vaccines, therapeutic antibodies, and drugs. Using a mammalian cell expression system, a genetically engineered sensor of fluorescent protein (Gamillus)-fused SARS-CoV-2 spike trimer (STG) to probe the viral entry process is developed. In ACE2-expressing cells, it is found that the STG probe has excellent performance in the live-cell visualization of receptor binding, cellular uptake, and intracellular trafficking of SARS-CoV-2 under virus-free conditions. The new system allows quantitative analyses of the inhibition potentials and detailed influence of COVID-19-convalescent human plasmas, neutralizing antibodies and compounds, providing a versatile tool for high-throughput screening and phenotypic characterization of SARS-CoV-2 entry inhibitors. This approach may also be adapted to develop a viral entry visualization system for other viruses.

18.
Emerg Microbes Infect ; 9(1): 2105-2113, 2020 Dec.
Article in English | MEDLINE | ID: covidwho-913100

ABSTRACT

The global pandemic of coronavirus disease 2019 (COVID-19) is a disaster for human society. A convenient and reliable neutralization assay is very important for the development of vaccines and novel drugs. In this study, a G protein-deficient vesicular stomatitis virus (VSVdG) bearing a truncated spike protein (S with C-terminal 18 amino acid truncation) was compared to that bearing the full-length spike protein of SARS-CoV-2 and showed much higher efficiency. A neutralization assay was established based on VSV-SARS-CoV-2-Sdel18 pseudovirus and hACE2-overexpressing BHK21 cells (BHK21-hACE2 cells). The experimental results can be obtained by automatically counting the number of EGFP-positive cells at 12 h after infection, making the assay convenient and high-throughput. The serum neutralizing titer measured by the VSV-SARS-CoV-2-Sdel18 pseudovirus assay has a good correlation with that measured by the wild type SARS-CoV-2 assay. Seven neutralizing monoclonal antibodies targeting the receptor binding domain (RBD) of the SARS-CoV-2 S protein were obtained. This efficient and reliable pseudovirus assay model could facilitate the development of new drugs and vaccines.


Subject(s)
Antibodies, Neutralizing/blood , Antibodies, Viral/blood , Betacoronavirus/immunology , Coronavirus Infections/diagnosis , Neutralization Tests/methods , Pneumonia, Viral/diagnosis , Spike Glycoprotein, Coronavirus/immunology , Animals , Antibodies, Monoclonal/immunology , Antibodies, Neutralizing/immunology , Antibodies, Viral/immunology , COVID-19 , Cell Line , Chlorocebus aethiops , Cricetinae , Pandemics , SARS-CoV-2 , Spike Glycoprotein, Coronavirus/genetics , Vero Cells , Vesicular stomatitis Indiana virus/genetics , Vesicular stomatitis Indiana virus/immunology
19.
medRxiv ; 2020 Oct 20.
Article in English | MEDLINE | ID: covidwho-900759

ABSTRACT

Since its first emergence from China in late 2019, the SARS-CoV-2 virus has spread globally despite unprecedented containment efforts, resulting in a catastrophic worldwide pandemic. Successful identification and isolation of infected individuals can drastically curtail virus spread and limit outbreaks. However, during the early stages of global transmission, point-of-care diagnostics were largely unavailable and continue to remain difficult to procure, greatly inhibiting public health efforts to mitigate spread. Furthermore, the most prevalent testing kits rely on reagent- and time-intensive protocols to detect viral RNA, preventing rapid and cost-effective diagnosis. Therefore the development of an extensive toolkit for point-of-care diagnostics that is expeditiously adaptable to new emerging pathogens is of critical public health importance. Recently, a number of novel CRISPR-based diagnostics have been developed to detect COVID-19. Herein, we outline the development of a CRISPR-based nucleic acid molecular diagnostic utilizing a Cas13d ribonuclease derived from Ruminococcus flavefaciens (CasRx) to detect SARS-CoV-2, an approach we term SENSR (Sensitive Enzymatic Nucleic-acid Sequence Reporter). We demonstrate SENSR robustly detects SARS-CoV-2 sequences in both synthetic and patient-derived samples by lateral flow and fluorescence, thus expanding the available point-of-care diagnostics to combat current and future pandemics.

20.
Preprint | SSRN | ID: ppcovidwho-738

ABSTRACT

Background: This study aimed to establish an effective diagnostic nomogram for suspected SARS-CoV-2 pneumonia patients. br br Methods: We used the LASSO aggre

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