Coronavirus SARS-CoV-2 Detection and Assay Development
O surgimento de novas doenças virais, incluindo os surtos de coronavírus SARS-CoV-2 (COVID 19), MERS (2015) e SARS (2003), destacam a importância de métodos rápidos para detectar e identificar os vírus alvo em grande escala. Eles também ilustram como os esforços de pesquisa para desenvolver medicamentos ou vacinas são acelerados em tempos de necessidade urgente. Durante o atual surto de coronavírus iniciado em 2019, a necessidade de fornecimentos e apoio aos cientistas que trabalham para reduzir a propagação do vírus é imediata.
A Promega apoia os cientistas que trabalham para compreender os mecanismos moleculares pelos quais novos vírus infectam humanos e animais, e para desenvolver métodos de detecção precisos. Nós fornecemos suporte técnico colaborativo, experiência em purificação de ácidos nucleicos, produtos de amplificação e tecnologias de rotulagem que agilizam os fluxos de trabalho e facilitam o trabalho dos laboratórios de pesquisa que estudam o coronavírus e outras doenças virais emergentes.
Produtos de Amplificação para Análise Viral
qPCR e RT-qPCR
Reagentes de PCR, incluindo produtos de qPCR e RT-qPCR, são usados para detectar e amplificar sequências-alvo virais. Estes reagentes são amplamente utilizados em estudos de pesquisa virológica.
PCR Reagents and Accessory Products
GoTaq® Probe qPCR and RT-qPCR Systems
Misturas principais prontas para uso para qPCR com base em sonda ou RT-qPCR.
A6101, A6102, A6120, A6121, A6110
XpressAmp® Direct Amplification Reagents
Fast, RNA extraction-free method to prepare viral samples for PCR-based amplification.
A8882, A8880
SARS-CoV-2 RT-qPCR Kit for Wastewater
Detect SARS-CoV-2 RNA in preprocessed wastewater with high efficiency and sensitivity.
AM2100, AM2110, AM2120, AM2130, AM2140, AM2150, AM2160, AM2170, AM2180, AM2070, AM2050, AM2060
Amplification Application Notes
Direct Amplification from Saliva
Viral RNA detection from Saliva using XpressAmp® lysates: GoTaq® Probe 1-Step RT-qPCR System; Luna® Universal Probe One-Step qPCR Kit
Viral RNA detection from Saliva using SalivaDirect™ and GoTaq® Probe 1-Step RT-qPCR System: CFX; 7500
Direct Amplification from Viral Transport Media
Viral RNA from VTM using XpressAmp® lysates: GoTaq® 1-Step RT-qPCR System
Referências
RT-PCR:
Diagne, M. et al. (2020) Dianke virus: A new mesonivirus species isolated from mosquitoes in Eastern Senegal. Virus Research 275, 2 January 2020, 197802.
Alves, M.T. et al. (2018) Mamastrovirus 5 detected in a crab-eating fox (Cerdocyon thous): Expanding wildlife host range of astroviruses. Comparative Immunology, Microbiology and Infectious Diseases 58, 36-43.
qPCR for Detection of Viral RNA:
Cai, M. et al. (2020) The R251K Substitution in Viral Protein PB2 Increases Viral Replication and Pathogenicity of Eurasian Avian-like H1N1 Swine Influenza Viruses. Viruses 12(1), 52.
Viral Structure and Characterization:
Wicht, O. et al. (2014) Proteolytic Activation of the Porcine Epidemic Diarrhea Coronavirus Spike Fusion Protein by Trypsin in Cell Culture. J. Virol. 88(14), 7952–7961.
Scalable Viral Nucleic Acid Extraction Solutions
Trusted Promega chemistries for viral nucleic acid purification are available in manual and automated formats to accommodate differing sample processing needs.
In addition to Maxwell® Instruments and cartridge-based kits for automated purification of 1-48 samples, Maxwell® HT extraction chemistries are available in plate-based, 96-well formats that can be used with high-throughput liquid-handling robots or performed manually with multichannel pipettors. We also offer a variety of viral manual extraction kits and protocols. These viral nucleic acid extraction chemistries are available to help laboratories developing coronavirus (SARS-CoV-2) tests and researching solutions for the COVID-19 pandemic.
Automated Viral Nucleic Acid Extraction
Maxwell® instruments are used together with Maxwell® Viral Total Nucleic Acid Purification Kits to extract viral nucleic acids from up to 48 samples per run.
Kits de Purificação de Ácido Nucleico Maxwell® Viral Total
Maxwell® RSC Viral Total Nucleic Acid Purification Kit
Viral total nucleic acid extraction from various samples types using the Maxwell® RSC Instrument.
AS1330, ASB1330
Viral RNA/DNA Concentration and Extraction Kits for Wastewater
Reagents and consumables to concentrate and purify total nucleic acids (TNA) in one convenient kit.
A2991, AS1831, AX9190, A3050, A3060, A3070, A3080
High-Throughput Automation and Manual Purification Chemistries
ReliaPrep™ Viral TNA Miniprep Kit, Custom
The ReliaPrep™ Viral TNA Miniprep Kit, available as a custom order, uses spin columns for rapid centrifugation-based processing of viral samples. See the Application Notes provided below for example protocols.
Maxwell® HT Viral TNA Kit, Custom
The Maxwell® HT Viral TNA Kit uses magnetic resin for easy automation and separation using robotic liquid handlers. The kit can also be used for manual purification in multiwell plates. For example protocols using the Maxwell® HT Viral TNA Kit on Tecan, Hamilton, Eppendorf and Kingfisher robots, and for example manual purification protocols, please review the Application Notes provided below.
Please contact us for information on pricing and availability.
Purification Scale-Up and Automation Support
Promega Field Support Scientists (FSS) are available to help labs implement or optimize automation of viral RNA extraction methods, regardless of the instrumentation platform used.
If you are new to automation, pressed for time, or need help choosing and adapting a viral nucleic acid purification method, we can help. We provide chemistry and instrumentation expertise and will work with you to configure your instrument and optimize the workflow to meet your laboratory's needs.
Viral RNA Extraction Application Notes
Manual Purification
ReliaPrep™ Viral TNA Miniprep Kit, Custom
Viral RNA from Transport Medium
Maxwell® HT Viral TNA Kit
Viral RNA from Sputum (96-well)
Maxwell® Instrument Purification
Maxwell® RSC Instrument
Viral RNA: CDC 2019-nCoV EUA Extraction Protocol
Viral RNA from Saliva: Oragene-RE-100; Oragene OM-505; Oragene OR-100; MP+ R-2000; Norgen RNA; Spectrum SDNA-1000
RNA from Viral Transport Medium
Viral RNA from Wastewater Concentrate
Maxwell® 16 Instrument
Referências
Extração de RNA Viral usando Maxwell® 48:
Jaeger, A.S. et al. (2018) Zika viruses of both African and Asian lineages cause fetal harm in a vertical transmission model. PLOS Neglected Tropical Diseases doi: 10.1371/journal. pntd.0007343
Chantal Reusken, C. et al. (2019) Toscana, West Nile, Usutu and tick-borne encephalitis viruses: external quality assessment for molecular detection of emerging neurotropic viruses in Europe, 2017. Euro Surveill. Dec 12; 24(50): 1900051.
Estamos comprometidos a apoiar nossos clientes que estão trabalhando para ajudar a encontrar soluções para as pessoas mais afetadas.
Tecnologias Repórteres para Pesquisa em Virologia
Luciferase Reporters
Repórteres baseados em Luciferase proporcionam um método simples e conveniente para monitorar os efeitos de tratamentos em potencial, criando ensaios in vitro para detectar anticorpos virais, e para imagiologia em modelos animais. O tamanho reduzido da NanoLuc® Luciferase faz com que ela seja apropriada para a criação de vírus repórteres.
Remdesivir (RDV ou GS-5734) foi usado no tratamento do primeiro caso de SARS-CoV-2 (2019-nCoV ) nos EUA. O Remdesivir ainda não é um medicamento aprovado em nenhum país, mas foi solicitado por várias agências de saúde pelo mundo para ajudar a combater a SARS-CoV-2. Você pode ler mais sobre o uso de repórteres NanoLuc® Luciferase para apoiar essas pesquisas no nosso blog, clicando aqui..
Referências
Evaluation of anti-viral drug candidates:
Zhang, L. et al. (2020) Alpha-ketoamides as broad-spectrum inhibitors of coronavirus and enterovirus replication. Structure-based design, synthesis, and activity assessment. J. Med Chem. DOI: 10.1021/ acs.jmedchem.9b01828. Publication Date (Web): 11 Feb 2020
Sheahan, T.P. et al. (2020) Comparative therapeutic efficacy of remdesivir and combination lopinavir, ritonavir, and interferon beta against MERS-CoV. Nature Communications 11, Article number: 222.
Testing for viral antibodies in serum using viral proteins fused to NanoLuc® luciferase:
Wang, W. et al. (2019) A novel luciferase immunosorbent assay performs better than a commercial enzyme-linked immunosorbent assay to detect MERS-CoV specific IgG in humans and animals. Biosafety and Health 1(3), 134–43.
Monitoring viral distribution by bioluminescence imaging:
Shen, L. et al. (2019) High-Throughput Screening and Identification of Potent Broad-Spectrum Inhibitors of Coronaviruses. DOI: 10.1128/ JVI.00023-19. J. Virol. 93(12), e00023-19.
Interessado em Vírus Repórteres NanoLuc®?
Reporter and cell health assays, RT-qPCR, and RNA production systems are widely used by researchers studying key aspects of viral pathogenesis, including characterizing virus:host interactions, tracking viral nucleic acids and proteins in the cell, and monitoring host cell viability and metabolism.
Serology and PCR Testing
Development and implementation of accurate molecular and serological assays is an urgent focus of many laboratories worldwide. Promega provides products and custom services to labs designing, performing and scaling up PCR-based molecular tests, and to support labs performing serological assays and researching new serological testing methods.
Coronavirus SARS-CoV-2 News and Updates
Last updated , Dec 23 2021
O Novo Coronavírus de 2019 (2019-nCoV) é um novo vírus que surgiu na China e tem estado nas manchetes desde o mês passado. Este vírus representa um grande desafio para pesquisadores e autoridades de saúde pública. Enquanto monitoramos o progresso da pesquisa global, estamos preparados para apoiar os cientistas em seu trabalho para compreender este vírus emergente e desenvolver medicamentos para tratar a infecção.
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