Viruses

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    Protocols in Current Issue
    COVID-19 Sample Pooling: From RNA Extraction to Quantitative Real-time RT-PCR

    The COVID-19 pandemic requires mass screening to identify those infected for isolation and quarantine. Individually screening large populations for the novel pathogen, Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), is costly and

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    COVID-19 Sample Pooling: From RNA Extraction to Quantitative Real-time RT-PCR
    [Abstract]

    The COVID-19 pandemic requires mass screening to identify those infected for isolation and quarantine. Individually screening large populations for the novel pathogen, Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), is costly and requires a lot of resources. Sample pooling methods improve the efficiency of mass screening and consume

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    Computational Analysis and Phylogenetic Clustering of SARS-CoV-2 Genomes
    Authors:  Bani Jolly and Vinod Scaria, date: 04/20/2021, view: 934, Q&A: 0
    [Abstract]

    COVID-19, the disease caused by the novel SARS-CoV-2 coronavirus, originated as an isolated outbreak in the Hubei province of China but soon created a global pandemic and is now a major threat to healthcare systems worldwide. Following the rapid human-to-human transmission of the infection, institutes around the world have made efforts to generate

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    Screening for Lysogen Activity in Therapeutically Relevant Bacteriophages
    Authors:  Fernando L. Gordillo Altamirano and Jeremy J. Barr, date: 04/20/2021, view: 833, Q&A: 1
    [Abstract]

    Lysogenic phages can integrate into their bacterial host’s genome, potentially transferring any genetic information they possess including virulence or resistance genes, and are therefore routinely excluded from therapeutic applications. Lysogenic behavior is typically seen in phages that create turbid plaques or possess subpar bactericidal

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    A Sensitive and Specific PCR-based Assay to Quantify Hepatitis B Virus Covalently Closed Circular (ccc) DNA while Preserving Cellular DNA
    Authors:  Benno Zehnder, Stephan Urban and Thomas Tu, date: 04/20/2021, view: 572, Q&A: 0
    [Abstract]

    Hepatitis B virus (HBV) is the major cause of liver diseases and liver cancer worldwide. After infecting hepatocytes, the virus establishes a stable episome (covalently closed circular DNA, or cccDNA) that serves as the template for all viral transcripts. Specific and accurate quantification of cccDNA is difficult because infected cells contain

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    Detection and Quantification of African Swine Fever Virus in MA-104 Cells
    [Abstract]

    Detection of live African swine fever virus (ASFV) has historically relied on the use of primary swine macrophages (PSM). PSM do not replicate and have to be isolated fresh from donor swine. We previously identified that a MA-104 cells (ATCC #CRL-2378.1), a commercially available cell line isolated from African green monkey (Cercopithecus

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    Generation and Implementation of Reporter BHK-21 Cells for Live Imaging of Flavivirus Infection
    Authors:  Jorge L. Arias-Arias and Rodrigo Mora-Rodríguez, date: 03/05/2021, view: 545, Q&A: 0
    [Abstract]

    The genus Flavivirus within the family Flaviviridae includes many viral species of medical importance, such as yellow fever virus (YFV), Zika virus (ZIKV), and dengue virus (DENV), among others. Presently, the identification of flavivirus-infected cells is based on either the immunolabeling of viral proteins, the application of recombinant

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    Primer ID Next-Generation Sequencing for the Analysis of a Broad Spectrum Antiviral Induced Transition Mutations and Errors Rates in a Coronavirus Genome
    [Abstract]

    Next generations sequencing (NGS) has become an important tool in biomedical research. The Primer ID approach combined with the MiSeq platform overcomes the limitation of PCR errors and reveals the true sampling depth of population sequencing, making it an ideal tool to study mutagenic effects of potential broad-spectrum antivirals on RNA viruses.

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    A One-enzyme RT-qPCR Assay for SARS-CoV-2, and Procedures for Reagent Production
    Authors:  Sanchita Bhadra, Andre C. Maranhao, Inyup Paik and Andrew D. Ellington, date: 01/20/2021, view: 659, Q&A: 1
    [Abstract]

    Given the scale of the ongoing COVID-19 pandemic, the need for reliable, scalable testing, and the likelihood of reagent shortages, especially in resource-poor settings, we have developed an RT-qPCR assay that relies on an alternative to conventional viral reverse transcriptases, a thermostable reverse transcriptase/DNA polymerase (RTX) (Ellefson

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    Evaluation of the Sequence Variability within the PCR Primer/Probe Target Regions of the SARS-CoV-2 Genome
    Authors:  Kashif Aziz Khan and Peter Cheung, date: 12/20/2020, view: 1429, Q&A: 0
    [Abstract] Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2; initially named 2019-nCoV) is responsible for the recent coronavirus disease (COVID-19) pandemic, and polymerase chain reaction (PCR) is the current standard method for diagnosis from patient samples. As PCR assays are prone to sequence mismatches due to mutations in the viral genome, it ...
    Giant Mimiviridae CsCl Purification Protocol
    Authors:  Lionel Bertaux, Audrey Lartigue and Sandra Jeudy, date: 11/20/2020, view: 809, Q&A: 0
    [Abstract] While different giant viruses’ purification protocols are available, they are not fully described and they use sucrose gradient that does not reach an equilibrium. Here, we report a protocol for the purification of members of the Mimiviridae family virions resulting from Acanthamoeaba castellanii infections. Viruses are ...



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