Microbiology

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    Protocols in Current Issue
    VirScan: High-throughput Profiling of Antiviral Antibody Epitopes
    Authors:  Ellen L. Shrock, Christine L. Shrock and Stephen J. Elledge, date: 07/05/2022, view: 1353, Q&A: 0
    [Abstract]

    Profiling the specificities of antibodies can reveal a wealth of information about humoral immune responses and the antigens they target. Here, we present a protocol for VirScan, an application of the phage immunoprecipitation sequencing (PhIP-Seq) method for profiling the specificities of human antiviral antibodies. Accompanying this protocol is

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    Ex vivo Human Skin Infection with Herpes Simplex Virus 1
    Authors:  Nydia C. De La Cruz, Maureen Möckel, Lisa Wirtz and Dagmar Knebel-Mörsdorf, date: 05/05/2022, view: 722, Q&A: 0
    [Abstract]

    Although herpes simplex virus 1 (HSV-1) is a well-studied virus, how the virus invades its human host via skin and mucosa to reach its receptors and initiate infection remains an open question. For studies of HSV-1 infection in skin, mice have been used as animal models. Murine skin infection can be induced after injection or scratching of the

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    Production of Recombinant Hepatitis B virus (HBV) and Detection of HBV in Infected Human Liver Organoids
    Authors:  Tanvir Hossain, Shahla Romal and Tokameh Mahmoudi, date: 04/20/2022, view: 894, Q&A: 0
    [Abstract]

    The absence of long term, primary untransformed in vitro models that support hepatitis B virus (HBV) infection and replication have hampered HBV pre-clinical research, which was reflected in the absence of a curative therapy until recently. One of the limitations for in vitro HBV research has been the absence of high titer and pure recombinant HBV

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    Generation, Maintenance and HBV Infection of Human Liver Organoids
    Authors:  Shahla Romal, Tanvir Hossain and Tokameh Mahmoudi, date: 03/20/2022, view: 1454, Q&A: 0
    [Abstract]

    Hepatitis B virus (HBV) infection represents a major public health problem infecting approximately 400 million people worldwide. Despite the availability of a preventive vaccine and anti-viral therapies, chronic HBV infection remains a major health issue because it increases the risk of developing liver cirrhosis and hepatocellular carcinoma

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    Three-dimensional Models of the Nasopharynx for the Study of Epstein-Barr Virus Infection
    [Abstract]

    The ubiquitous and cancer-associated Epstein-Barr virus (EBV) is associated with nearly all cases of nasopharyngeal carcinoma (NPC). Nasopharyngeal tissue is comprised of both pseudostratified and stratified epithelium, which are modeled in three-dimensional (3-D) cell culture. The cellular origin of EBV-associated NPC is as yet unknown, but both

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    Amber Suppression Technology for Mapping Site-specific Viral-host Protein Interactions in Mammalian Cells
    Authors:  Nur Firdaus Isa, Olivier Bensaude and Shona Murphy, date: 02/05/2022, view: 1641, Q&A: 0
    [Abstract]

    Probing the molecular interactions of viral-host protein complexes to understand pathogenicity is essential in modern virology to help the development of antiviral therapies. Common binding assays, such as co-immunoprecipitation or pull-downs, are helpful in investigating intricate viral-host proteins interactions. However, such assays may miss

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    Intrathoracic Inoculation of Zika Virus in Aedes aegypti
    Authors:  Irma Sanchez-Vargas, Adeline E. Williams, Alexander W.E. Franz and Ken E. Olson, date: 09/20/2021, view: 1377, Q&A: 0
    [Abstract]

    Aedes aegypti mosquitoes are the main vectors of many medically relevant arthropod-borne (arbo) viruses, including Zika (ZIKV), dengue (DENV), and yellow fever (YFV). Vector competence studies with Ae. aegypti often involve challenging mosquitoes with an artificial bloodmeal containing virus and later quantifying viral titer or infectious

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    Optimised Method for the Production and Titration of Lentiviral Vectors Pseudotyped with the SARS-CoV-2 Spike
    [Abstract]

    The use of recombinant lentivirus pseudotyped with the coronavirus Spike protein of SARS-CoV-2 would circumvent the requirement of biosafety-level 3 (BSL-3) containment facilities for the handling of SARS-CoV-2 viruses. Herein, we describe a fast and reliable protocol for the transient production of lentiviruses pseudotyped with SARS-CoV-2 Spike

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    Visualization of Host Cell Kinase Activation by Viral Proteins Using GFP Fluorescence Complementation and Immunofluorescence Microscopy
    Authors:  Sherry T. Shu, Wing Fai Li and Thomas E. Smithgall, date: 07/05/2021, view: 3145, Q&A: 0
    [Abstract]

    Non-receptor protein-tyrosine kinases regulate cellular responses to many external signals and are important drug discovery targets for cancer and infectious diseases. While many assays exist for the assessment of kinase activity in vitro, methods that report changes in tyrosine kinase activity in single cells have the potential to provide

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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: 4273, 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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