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1.
Biosens Bioelectron ; 171: 112715, 2021 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-33099241

RESUMO

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes coronavirus disease 2019 (COVID-19), a newly emerging human infectious disease. Because no specific antiviral drugs or vaccines are available to treat COVID-19, early diagnostics, isolation, and prevention are crucial for containing the outbreak. Molecular diagnostics using reverse transcription polymerase chain reaction (RT-PCR) are the current gold standard for detection. However, viral RNAs are much less stable during transport and storage than proteins such as antigens and antibodies. Consequently, false-negative RT-PCR results can occur due to inadequate collection of clinical specimens or poor handling of a specimen during testing. Although antigen immunoassays are stable diagnostics for detection of past infection, infection progress, and transmission dynamics, no matched antibody pair for immunoassay of SARS-CoV-2 antigens has yet been reported. In this study, we designed and developed a novel rapid detection method for SARS-CoV-2 spike 1 (S1) protein using the SARS-CoV-2 receptor ACE2, which can form matched pairs with commercially available antibodies. ACE2 and S1-mAb were paired with each other for capture and detection in a lateral flow immunoassay (LFIA) that did not cross-react with SARS-CoV Spike 1 or MERS-CoV Spike 1 protein. The SARS-CoV-2 S1 (<5 ng of recombinant proteins/reaction) was detected by the ACE2-based LFIA. The limit of detection of our ACE2-LFIA was 1.86 × 105 copies/mL in the clinical specimen of COVID-19 Patients without no cross-reactivity for nasal swabs from healthy subjects. This is the first study to detect SARS-CoV-2 S1 antigen using an LFIA with matched pair consisting of ACE2 and antibody. Our findings will be helpful to detect the S1 antigen of SARS-CoV-2 from COVID-19 patients.


Assuntos
Betacoronavirus/isolamento & purificação , Técnicas Biossensoriais/instrumentação , Técnicas de Laboratório Clínico , Infecções por Coronavirus/diagnóstico , Peptidil Dipeptidase A/química , Pneumonia Viral/diagnóstico , Glicoproteína da Espícula de Coronavírus/análise , Anticorpos Monoclonais/química , Técnicas Biossensoriais/economia , Técnicas de Laboratório Clínico/economia , Técnicas de Laboratório Clínico/instrumentação , Infecções por Coronavirus/economia , Desenho de Equipamento , Humanos , Imunoensaio/economia , Imunoensaio/instrumentação , Imunoconjugados/química , Pandemias , Sensibilidade e Especificidade , Fatores de Tempo
2.
Talanta ; 222: 121534, 2021 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-33167242

RESUMO

As COVID-19 has reached pandemic status and the number of cases continues to grow, widespread availability of diagnostic testing is critical in helping identify and control the emergence of this rapidly spreading and serious illness. However, a lacking in making a quick reaction to the threat and starting early development of diagnostic sensing tools has had an important impact globally. In this regard, here we will review critically the current developed diagnostic tools in response to the COVID-19 pandemic and compare the different types through the discussion of their pros and cons such as nucleic acid detection tests (including PCR and CRISPR), antibody and protein-based diagnosis tests. In addition, potential technologies that are under development such as on-site diagnosis platforms, lateral flow, and portable PCR units are discussed. Data collection and epidemiological analysis could also be an interesting factor to incorporate with the emerging technologies especially with the wide access to smartphones. Lastly, a SWOT analysis and perspectives on how the development of novel sensory platforms should be treated by the different decision-makers are analyzed.


Assuntos
Técnicas de Laboratório Clínico/métodos , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/epidemiologia , Pandemias , Pneumonia Viral/diagnóstico , Pneumonia Viral/epidemiologia , Técnicas de Laboratório Clínico/instrumentação , Humanos , Testes Imediatos , Kit de Reagentes para Diagnóstico
3.
Ann Biol Clin (Paris) ; 78(6): 609-616, 2020 Dec 01.
Artigo em Francês | MEDLINE | ID: mdl-33361015

RESUMO

Confronted with the COVID-19 crisis, healthcare professionals have had to tackle an epidemic crisis of a huge magnitude for which they were not prepared. Medical laboratories have been on the front line, from collecting samples to performing the analysis required to diagnose this new pathology. Responding to the needs and to the urgency of the situation, the authorities relied on the network of private laboratories. In France, private laboratory medicine represents 70% of overall activity, and with a network of more than 4,000 local laboratories, private laboratory medicine has been the cornerstone of the « screen-trace-isolate ¼ strategy. This article gives feedback from private laboratory medicine professionals, directly involved in the reorganization carried out at the pre-analytical, analytical and post-analytical stages, during the crisis from March to October 2020.


Assuntos
/epidemiologia , Serviços de Laboratório Clínico/organização & administração , Pandemias , Setor Privado/organização & administração , Manejo de Espécimes/normas , /diagnóstico , Serviços de Laboratório Clínico/normas , Técnicas de Laboratório Clínico/instrumentação , Técnicas de Laboratório Clínico/métodos , Técnicas de Laboratório Clínico/normas , Infecção Hospitalar/epidemiologia , Infecção Hospitalar/prevenção & controle , Segurança de Equipamentos/métodos , Segurança de Equipamentos/normas , França/epidemiologia , Unidades Hospitalares/organização & administração , Humanos , Colaboração Intersetorial , Corpo Clínico/organização & administração , Corpo Clínico/normas , Segurança do Paciente/normas , Fase Pré-Analítica/métodos , Fase Pré-Analítica/normas , Setor Privado/normas , Manejo de Espécimes/métodos
5.
Comput Math Methods Med ; 2020: 5248569, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33082839

RESUMO

In this paper, we developed a model that suggests the use of robots in identifying COVID-19-positive patients and which studied the effectiveness of the government policy of prohibiting migration of individuals into their countries especially from those countries that were known to have COVID-19 epidemic. Two compartmental models consisting of two equations each were constructed. The models studied the use of robots for the identification of COVID-19-positive patients. The effect of migration ban strategy was also studied. Four biologically meaningful equilibrium points were found. Their local stability analysis was also carried out. Numerical simulations were carried out, and the most effective strategy to curtail the spread of the disease was shown.


Assuntos
Betacoronavirus , Infecções por Coronavirus/prevenção & controle , Modelos Biológicos , Pandemias/prevenção & controle , Pneumonia Viral/prevenção & controle , Técnicas de Laboratório Clínico/instrumentação , Técnicas de Laboratório Clínico/estatística & dados numéricos , Biologia Computacional , Simulação por Computador , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/transmissão , Humanos , Conceitos Matemáticos , Modelos Estatísticos , Pandemias/estatística & dados numéricos , Pneumonia Viral/epidemiologia , Pneumonia Viral/transmissão , Robótica/instrumentação , Robótica/estatística & dados numéricos , Viagem
7.
Biosens Bioelectron ; 170: 112673, 2020 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-33038584

RESUMO

Currently the world is being challenged by a public health emergency caused by the coronavirus pandemic (COVID-19). Extensive efforts in testing for coronavirus infection, combined with isolating infected cases and quarantining those in contact, have proven successful in bringing the epidemic under control. Rapid and facile screening of this disease is in high demand. This review summarises recent advances in strategies reported by international researchers and engineers concerning how to tackle COVID-19 via rapid testing, mainly through nucleic acid- and antibody- testing. The roles of biosensors as powerful analytical tools are emphasized for the detection of viral RNAs, surface antigens, whole viral particles, antibodies and other potential biomarkers in human specimen. We critically review in depth newly developed biosensing methods especially for in-field and point-of-care detection of SARS-CoV-2. Additionally, this review describes possible future strategies for virus rapid detection. It helps researchers working on novel sensor technologies to tailor their technologies in a way to address the challenge for effective detection of COVID-19.


Assuntos
Betacoronavirus/isolamento & purificação , Técnicas Biossensoriais , Técnicas de Laboratório Clínico , Infecções por Coronavirus/diagnóstico , Pneumonia Viral/diagnóstico , Testes Imediatos , Animais , Técnicas Biossensoriais/economia , Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/métodos , Técnicas de Laboratório Clínico/economia , Técnicas de Laboratório Clínico/instrumentação , Técnicas de Laboratório Clínico/métodos , Infecções por Coronavirus/economia , Desenho de Equipamento , Humanos , Pandemias , Testes Imediatos/economia , Fatores de Tempo
8.
IEEE Pulse ; 11(5): 28-31, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33064642

RESUMO

An at-home test for coronavirus disease 2019 (COVID-19) could be released commercially as early as August, according to Scanwell Health of Los Angeles. A combination of a finger-prick blood sample and a smart-phone app, the test is designed to detect the presence of antibodies to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus that causes COVID-19. The company hopes to receive Emergency Use Authorization from the U.S. Food and Drug Administration (FDA) by the end of summer, and make its first commercially available tests soon after.


Assuntos
Anticorpos Antivirais/sangue , Betacoronavirus/imunologia , Técnicas de Laboratório Clínico/métodos , Infecções por Coronavirus/diagnóstico , Pneumonia Viral/diagnóstico , Técnicas de Laboratório Clínico/instrumentação , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/imunologia , Humanos , Aplicativos Móveis , Pandemias , Participação do Paciente/métodos , Pneumonia Viral/epidemiologia , Pneumonia Viral/imunologia , Estados Unidos/epidemiologia
9.
Biosens Bioelectron ; 166: 112431, 2020 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-32862842

RESUMO

Last few decades, viruses are a real menace to human safety. Therefore, the rapid identification of viruses should be one of the best ways to prevent an outbreak and important implications for medical healthcare. The recent outbreak of coronavirus disease (COVID-19) is an infectious disease caused by a newly discovered coronavirus which belongs to the single-stranded, positive-strand RNA viruses. The pandemic dimension spread of COVID-19 poses a severe threat to the health and lives of seven billion people worldwide. There is a growing urgency worldwide to establish a point-of-care device for the rapid detection of COVID-19 to prevent subsequent secondary spread. Therefore, the need for sensitive, selective, and rapid diagnostic devices plays a vital role in selecting appropriate treatments and to prevent the epidemics. During the last decade, electrochemical biosensors have emerged as reliable analytical devices and represent a new promising tool for the detection of different pathogenic viruses. This review summarizes the state of the art of different virus detection with currently available electrochemical detection methods. Moreover, this review discusses different fabrication techniques, detection principles, and applications of various virus biosensors. Future research also looks at the use of electrochemical biosensors regarding a potential detection kit for the rapid identification of the COVID-19.


Assuntos
Betacoronavirus , Técnicas Biossensoriais/instrumentação , Técnicas de Laboratório Clínico/instrumentação , Infecções por Coronavirus/diagnóstico , Técnicas Eletroquímicas/instrumentação , Pneumonia Viral/diagnóstico , Vírus/isolamento & purificação , Animais , Betacoronavirus/isolamento & purificação , Betacoronavirus/patogenicidade , Infecções por Coronavirus/virologia , Desenho de Equipamento , Humanos , Nanopartículas Metálicas/química , Nanopartículas Metálicas/ultraestrutura , Microscopia Eletrônica de Varredura , Pandemias , Pneumonia Viral/virologia , Testes Imediatos , Vírus/patogenicidade
10.
Nat Commun ; 11(1): 4464, 2020 09 08.
Artigo em Inglês | MEDLINE | ID: mdl-32900994

RESUMO

The SARS-CoV-2 pandemic has shown how a rapid rise in demand for patient and community sample testing can quickly overwhelm testing capability globally. With most diagnostic infrastructure dependent on specialized instruments, their exclusive reagent supplies quickly become bottlenecks, creating an urgent need for approaches to boost testing capacity. We address this challenge by refocusing the London Biofoundry onto the development of alternative testing pipelines. Here, we present a reagent-agnostic automated SARS-CoV-2 testing platform that can be quickly deployed and scaled. Using an in-house-generated, open-source, MS2-virus-like particle (VLP) SARS-CoV-2 standard, we validate RNA extraction and RT-qPCR workflows as well as two detection assays based on CRISPR-Cas13a and RT-loop-mediated isothermal amplification (RT-LAMP). In collaboration with an NHS diagnostic testing lab, we report the performance of the overall workflow and detection of SARS-CoV-2 in patient samples using RT-qPCR, CRISPR-Cas13a, and RT-LAMP. The validated RNA extraction and RT-qPCR platform has been installed in NHS diagnostic labs, increasing testing capacity by 1000 samples per day.


Assuntos
Betacoronavirus/genética , Técnicas de Laboratório Clínico/métodos , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/virologia , Pneumonia Viral/diagnóstico , Pneumonia Viral/virologia , Betacoronavirus/isolamento & purificação , Bioensaio , Sistemas CRISPR-Cas , Técnicas de Laboratório Clínico/instrumentação , Técnicas de Laboratório Clínico/normas , Humanos , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Pandemias , RNA Viral/análise , RNA Viral/genética , RNA Viral/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real , Sensibilidade e Especificidade
11.
Sensors (Basel) ; 20(18)2020 Sep 08.
Artigo em Inglês | MEDLINE | ID: mdl-32911861

RESUMO

Lung sounds acquired by stethoscopes are extensively used in diagnosing and differentiating respiratory diseases. Although an extensive know-how has been built to interpret these sounds and identify diseases associated with certain patterns, its effective use is limited to individual experience of practitioners. This user-dependency manifests itself as a factor impeding the digital transformation of this valuable diagnostic tool, which can improve patient outcomes by continuous long-term respiratory monitoring under real-life conditions. Particularly patients suffering from respiratory diseases with progressive nature, such as chronic obstructive pulmonary diseases, are expected to benefit from long-term monitoring. Recently, the COVID-19 pandemic has also shown the lack of respiratory monitoring systems which are ready to deploy in operational conditions while requiring minimal patient education. To address particularly the latter subject, in this article, we present a sound acquisition module which can be integrated into a dedicated garment; thus, minimizing the role of the patient for positioning the stethoscope and applying the appropriate pressure. We have implemented a diaphragm-less acousto-electric transducer by stacking a silicone rubber and a piezoelectric film to capture thoracic sounds with minimum attenuation. Furthermore, we benchmarked our device with an electronic stethoscope widely used in clinical practice to quantify its performance.


Assuntos
Betacoronavirus , Técnicas de Laboratório Clínico/instrumentação , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/fisiopatologia , Monitorização Ambulatorial/instrumentação , Pneumonia Viral/diagnóstico , Pneumonia Viral/fisiopatologia , Sons Respiratórios/diagnóstico , Sons Respiratórios/fisiopatologia , Estetoscópios , Dispositivos Eletrônicos Vestíveis , Acústica , Auscultação/instrumentação , Impedância Elétrica , Desenho de Equipamento , Humanos , Pandemias , Tecnologia de Sensoriamento Remoto/instrumentação , Processamento de Sinais Assistido por Computador , Transdutores , Tecnologia sem Fio/instrumentação
13.
Biosens Bioelectron ; 166: 112436, 2020 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-32750677

RESUMO

Our recent experience of the COVID-19 pandemic has highlighted the importance of easy-to-use, quick, cheap, sensitive and selective detection of virus pathogens for the efficient monitoring and treatment of virus diseases. Early detection of viruses provides essential information about possible efficient and targeted treatments, prolongs the therapeutic window and hence reduces morbidity. Graphene is a lightweight, chemically stable and conductive material that can be successfully utilized for the detection of various virus strains. The sensitivity and selectivity of graphene can be enhanced by its functionalization or combination with other materials. Introducing suitable functional groups and/or counterparts in the hybrid structure enables tuning of the optical and electrical properties, which is particularly attractive for rapid and easy-to-use virus detection. In this review, we cover all the different types of graphene-based sensors available for virus detection, including, e.g., photoluminescence and colorimetric sensors, and surface plasmon resonance biosensors. Various strategies of electrochemical detection of viruses based on, e.g., DNA hybridization or antigen-antibody interactions, are also discussed. We summarize the current state-of-the-art applications of graphene-based systems for sensing a variety of viruses, e.g., SARS-CoV-2, influenza, dengue fever, hepatitis C virus, HIV, rotavirus and Zika virus. General principles, mechanisms of action, advantages and drawbacks are presented to provide useful information for the further development and construction of advanced virus biosensors. We highlight that the unique and tunable physicochemical properties of graphene-based nanomaterials make them ideal candidates for engineering and miniaturization of biosensors.


Assuntos
Betacoronavirus/isolamento & purificação , Técnicas Biossensoriais , Técnicas de Laboratório Clínico , Infecções por Coronavirus/diagnóstico , Grafite , Pneumonia Viral/diagnóstico , Vírus/isolamento & purificação , Reações Antígeno-Anticorpo , Betacoronavirus/genética , Betacoronavirus/patogenicidade , Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/métodos , Técnicas Biossensoriais/tendências , Técnicas de Laboratório Clínico/instrumentação , Técnicas de Laboratório Clínico/métodos , Técnicas de Laboratório Clínico/estatística & dados numéricos , Colorimetria , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/virologia , DNA Viral/análise , DNA Viral/genética , Técnicas Eletroquímicas , Desenho de Equipamento , Grafite/química , Humanos , Luminescência , Nanoestruturas/química , Hibridização de Ácido Nucleico , Pandemias , Pneumonia Viral/epidemiologia , Pneumonia Viral/virologia , Pontos Quânticos/química , Análise Espectral Raman , Ressonância de Plasmônio de Superfície , Virologia/métodos , Vírus/genética , Vírus/patogenicidade
14.
Biosens Bioelectron ; 166: 112455, 2020 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-32739797

RESUMO

The rapid spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has led to the coronavirus disease 2019 (COVID-19) worldwide pandemic. This unprecedented situation has garnered worldwide attention. An effective strategy for controlling the COVID-19 pandemic is to develop highly accurate methods for the rapid identification and isolation of SARS-CoV-2 infected patients. Many companies and institutes are therefore striving to develop effective methods for the rapid detection of SARS-CoV-2 ribonucleic acid (RNA), antibodies, antigens, and the virus. In this review, we summarize the structure of the SARS-CoV-2 virus, its genome and gene expression characteristics, and the current progression of SARS-CoV-2 RNA, antibodies, antigens, and virus detection. Further, we discuss the reasons for the observed false-negative and false-positive RNA and antibody detection results in practical clinical applications. Finally, we provide a review of the biosensors which hold promising potential for point-of-care detection of COVID-19 patients. This review thereby provides general guidelines for both scientists in the biosensing research community and for those in the biosensor industry to develop a highly sensitive and accurate point-of-care COVID-19 detection system, which would be of enormous benefit for controlling the current COVID-19 pandemic.


Assuntos
Betacoronavirus/isolamento & purificação , Técnicas Biossensoriais/métodos , Técnicas de Laboratório Clínico/métodos , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/virologia , Pandemias , Pneumonia Viral/diagnóstico , Pneumonia Viral/virologia , Anticorpos Antivirais/isolamento & purificação , Antígenos Virais/isolamento & purificação , Betacoronavirus/genética , Betacoronavirus/imunologia , Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/tendências , Técnicas de Laboratório Clínico/instrumentação , Técnicas de Laboratório Clínico/tendências , Infecções por Coronavirus/epidemiologia , Desenho de Equipamento , Genoma Viral , Humanos , Técnicas de Diagnóstico Molecular , Técnicas de Amplificação de Ácido Nucleico , Pneumonia Viral/epidemiologia , Testes Imediatos , RNA Viral/genética , RNA Viral/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Vírion/isolamento & purificação
16.
Med J Aust ; 213(6): 276-279, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32772375

RESUMO

OBJECTIVES: To design and evaluate 3D-printed nasal swabs for collection of samples for SARS-CoV-2 testing. DESIGN: An iterative design process was employed. Laboratory evaluation included in vitro assessment of mock nasopharyngeal samples spiked with two different concentrations of gamma-irradiated SARS-CoV-2. A prospective clinical study compared SARS-CoV-2 and human cellular material recovery by 3D-printed swabs and standard nasopharyngeal swabs. SETTING, PARTICIPANTS: Royal Melbourne Hospital, May 2020. Participants in the clinical evaluation were 50 hospital staff members attending a COVID-19 screening clinic and two inpatients with laboratory-confirmed COVID-19. INTERVENTION: In the clinical evaluation, a flocked nasopharyngeal swab sample was collected with the Copan ESwab and a mid-nasal sample from the other nostril was collected with the 3D-printed swab. RESULTS: In the laboratory evaluation, qualitative agreement with regard to SARS-CoV-2 detection in mock samples collected with 3D-printed swabs and two standard swabs was complete. In the clinical evaluation, qualitative agreement with regard to RNase P detection (a surrogate measure of adequate collection of human cellular material) in samples collected from 50 hospital staff members with standard and 3D-printed swabs was complete. Qualitative agreement with regard to SARS-CoV-2 detection in three pairs of 3D-printed mid-nasal and standard swab samples from two inpatients with laboratory-confirmed SARS-CoV-2 was also complete. CONCLUSIONS: Using 3D-printed swabs to collect nasal samples for SARS-CoV-2 testing is feasible, acceptable to patients and health carers, and convenient.


Assuntos
Técnicas de Laboratório Clínico/instrumentação , Infecções por Coronavirus/diagnóstico , Técnicas de Diagnóstico do Sistema Respiratório/instrumentação , Aceitação pelo Paciente de Cuidados de Saúde/estatística & dados numéricos , Pneumonia Viral/diagnóstico , Impressão Tridimensional , Adulto , Betacoronavirus/isolamento & purificação , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Nasofaringe/virologia , Pandemias
17.
Skeletal Radiol ; 49(11): 1873-1877, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32827082

RESUMO

The ongoing coronavirus disease 2019 (COVID-19) pandemic has increased the need for safe and efficient testing as a key containment strategy. Drive-through testing with nasopharyngeal swab has been implemented in many places in the USA as it allows for expeditious testing of large numbers of patients, limits healthcare workers' risk of exposure, and minimizes the use of personal protective equipment. We present a case where the aluminum shaft of the nasopharyngeal swab fractured during specimen collection at a drive-through testing facility and was suspected to have remained in the asymptomatic patient. Initial evaluation with a series of radiographs covering the skull base, neck, chest, and abdomen did not reveal the swab. On further clinical evaluation, the swab was found endoscopically, lodged between the left inferior turbinate and nasal floor, and was removed by an otorhinolaryngologist. Using a phantom model, we aimed to delineate an imaging technique to better visualize the aluminum shaft of the nasopharyngeal swab on radiographs to help in identification. A technique using lower tube voltage (kVp) with tight collimation centered at the nasal bones area produced the best visualization of the aluminum shaft of the swab. Recognition that aluminum foreign bodies may be difficult to visualize radiographically and  optimization of radiograph acquisition technique may help guide clinical management in unusual cases. Further evaluation with computed tomography or endoscopy should be considered in suspected cases where radiographs are negative.


Assuntos
Betacoronavirus , Técnicas de Laboratório Clínico/instrumentação , Infecções por Coronavirus/diagnóstico , Falha de Equipamento , Corpos Estranhos/diagnóstico por imagem , Pneumonia Viral/diagnóstico , Manejo de Espécimes/instrumentação , Idoso de 80 Anos ou mais , Alumínio , Humanos , Masculino , Pandemias , Radiografia/métodos , Instrumentos Cirúrgicos
18.
Virus Res ; 288: 198129, 2020 10 15.
Artigo em Inglês | MEDLINE | ID: mdl-32822689

RESUMO

The COVID-19 pandemic caused by SARS-CoV-2 affects all aspects of human life. Detection platforms that are efficient, rapid, accurate, specific, sensitive, and user friendly are urgently needed to manage and control the spread of SARS-CoV-2. RT-qPCR based methods are the gold standard for SARS-CoV-2 detection. However, these methods require trained personnel, sophisticated infrastructure, and a long turnaround time, thereby limiting their usefulness. Reverse transcription-loop-mediated isothermal amplification (RT-LAMP), a one-step nucleic acid amplification method conducted at a single temperature, has been used for colorimetric virus detection. CRISPR-Cas12 and CRISPR-Cas13 systems, which possess collateral activity against ssDNA and RNA, respectively, have also been harnessed for virus detection. Here, we built an efficient, rapid, specific, sensitive, user-friendly SARS-CoV-2 detection module that combines the robust virus amplification of RT-LAMP with the specific detection ability of SARS-CoV-2 by CRISPR-Cas12. Furthermore, we combined the RT-LAMP-CRISPR-Cas12 module with lateral flow cells to enable highly efficient point-of-care SARS-CoV-2 detection. Our iSCAN SARS-CoV-2 detection module, which exhibits the critical features of a robust molecular diagnostic device, should facilitate the effective management and control of COVID-19.


Assuntos
Betacoronavirus/genética , Sistemas CRISPR-Cas , Técnicas de Laboratório Clínico/métodos , Colorimetria/métodos , Infecções por Coronavirus/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Pneumonia Viral/diagnóstico , Técnicas de Laboratório Clínico/instrumentação , Colorimetria/instrumentação , Infecções por Coronavirus/virologia , Endodesoxirribonucleases/química , Humanos , Técnicas de Diagnóstico Molecular/instrumentação , Técnicas de Amplificação de Ácido Nucleico/instrumentação , Pandemias , Pneumonia Viral/virologia , Sistemas Automatizados de Assistência Junto ao Leito , Reologia , Sensibilidade e Especificidade
19.
J Appl Lab Med ; 5(6): 1307-1312, 2020 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-32761092

RESUMO

BACKGROUND: Numerous nucleic acid amplification assays utilizing different target genes of the SARS-CoV-2 genome have received emergency use authorization (EUA) by the United States Food and Drug Administration (FDA). Limited data are available comparing the test performance characteristics of these assays. METHODS: A diagnostic comparison study was performed to evaluate the performance of the Cepheid Xpert Xpress SARS-CoV-2 assay compared to the Hologic Panther Fusion SARS-CoV-2 assay using clinical nasopharyngeal specimens. Agreement between the two assays was assessed by overall, positive, and negative percent agreement and Cohen's kappa coefficient. RESULTS: A total of 104 (54 positive and 50 negative) clinical nasopharyngeal samples were tested by both assays. Using the Panther Fusion as a reference standard, the Xpert demonstrated an overall agreement of 99.0% [95% confidence interval (CI): 94.8-100], positive percent agreement of 98.1% (95% CI: 90.1-100), and a negative percent agreement of 100% (95% CI: 94.2-100). The kappa coefficient was 0.98 (95% CI: 0.94-1.0). One sample positive by the Panther Fusion with a cycle threshold (Ct) of 38.6 was found to be reproducibly negative by the Xpert assay. CONCLUSIONS: The Cepheid Xpert Xpress SARS-CoV-2 assay provides test performance comparable to the Hologic Panther Fusion SARS-CoV-2 assay while offering laboratories rapid, on-demand testing capacity.


Assuntos
Betacoronavirus/isolamento & purificação , Técnicas de Laboratório Clínico/instrumentação , Infecções por Coronavirus/diagnóstico , Técnicas de Diagnóstico Molecular/instrumentação , Pneumonia Viral/diagnóstico , RNA Viral/isolamento & purificação , Automação Laboratorial/instrumentação , Betacoronavirus/genética , Técnicas de Laboratório Clínico/estatística & dados numéricos , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/virologia , Humanos , Técnicas de Diagnóstico Molecular/estatística & dados numéricos , Reação em Cadeia da Polimerase Multiplex/instrumentação , Pandemias , Pneumonia Viral/epidemiologia , Pneumonia Viral/virologia , Sistemas Automatizados de Assistência Junto ao Leito/estatística & dados numéricos , Kit de Reagentes para Diagnóstico/estatística & dados numéricos , Reação em Cadeia da Polimerase em Tempo Real/instrumentação , Reprodutibilidade dos Testes , Fatores de Tempo , Estados Unidos/epidemiologia
20.
Life Sci ; 258: 118207, 2020 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-32777301

RESUMO

Due to COVID 19 outbreak many studies are being conducted for therapeutic strategies and vaccines but detection methods play an important role in the containment of the disease. Hence, this systematic review aims to evaluate the effectiveness of the molecular detection techniques in COVID-19. For framing the systematic review 6 literature databases (PubMed, EMBASE, OVID, Web of Science, Scopus and Google Scholar) were searched for relevant studies and articles were screened for relevant content till 25th April 2020. Observations from this systematic review reveal the utility of RT-PCR with serological testing as one such method cannot correlate with accurate results. Availability of point of care devices do not conform to sensitivity and specificity in comparison to the conventional methods due to lack of clinical investigations. Pivotal aim of molecular and serological research is the development of detection methods that can support the clinical decision making of patients suspected with SARS-CoV-2. However, none of the methods were 100% sensitive and specific; hence additional studies are required to overcome the challenges addressed here. We hope that the present article with its observations and suggestions will assist the researchers to realize this vision in future.


Assuntos
Betacoronavirus/isolamento & purificação , Técnicas de Laboratório Clínico , Infecções por Coronavirus/diagnóstico , Pneumonia Viral/diagnóstico , Betacoronavirus/genética , Técnicas de Laboratório Clínico/instrumentação , Técnicas de Laboratório Clínico/métodos , Infecções por Coronavirus/sangue , Humanos , Pandemias , Pneumonia Viral/sangue , Testes Imediatos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/instrumentação , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Sensibilidade e Especificidade
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