OHSU

Virology

virus_virologyWelcome to the Molecular Virology Support Core (MVSC) website!

The MVSC is a research support core at the Oregon National Primate Research Center (ONPRC), Oregon Health & Science University (OHSU) West Campus. The mission of MVSC is to provide essential virology services, reagents, specialized expertise and training to investigators in order to advance a wide array of research, with a particular focus on non-human primates (NHP). These range from basic studies in viral pathogenesis and immunology to viral vaccines, gene transfer, and gene therapy.

The MVSC supports investigators through an array of specialized virology services and the provision of technical expertise.  Key focus areas are:

  1. Custom production and quality control of viral vectors, viral stocks, and antigens
  2. Sensitive viral diagnostic assays to study virus spread after injection, monitor viral tissue distribution, and assess viral antibodies
  3. Provision and development of key reagents and standardized assays

In addition, the MVSC offers user training in virology techniques, safe handling of infectious agents, and assists in maintaining proper compliance with local institutional biosafety requirements.

Core Services

Adenovirus    
•    For custom adenoviral vector production, user clones genetic sequences of choice into a provided shuttle plasmid 

•    Production size can be anywhere from a small pilot stock to a large prep, with purity levels varying from crude lysate to highly purified stock.

•    Transgene cassette choices available:

o    Inducible promoter for expression of toxic proteins
o    CMV promoter and V5-tag for high-level expression and easy protein detection
o    RNA interference (RNAi) cassettes for expression of short hairpin RNA (shRNA)
o    Empty expression cassettes for user-supplied regulatory elements and transgenes
o    Pre-made common stock vectors (such as Adeno-CMV-eGFP) available for pilot testing
  • Adenovirus titration (particle titer; TCID50)

•    Adenovirus replication-competent virus (RCV) assay (required for in vivo use)

Adeno-Associated Virus (AAV)

  • For custom AAV vector production, user clones genetic sequences of choice into a provided ITR shuttle plasmid
  • Standard AAV production sizes range from 2,000 to 6,000 cm2 culture scale with purification by iodixanol step gradient
  • Transgene cassette choices available:
    • CMV promoter for high-level expression
    • Various marker gene and promoter cassettes (e.g. CAG, ER1alpha, EGFP, EYFP, dsRed, mCherry, IoxP)
    • RNA interference (RNAi) cassettes for expression of short hairpin RNA (shRNA) along with eGFP tracer
    • Self-complementary/double-stranded backbone vectors
    • Empty expression cassettes for user-supplied regulatory elements and transgenes
    • Pre-made AAV-CMV-EGFP stock vectors of different serotypes available for pilot testing
  • Other assays available:
  • AAV vector titration (vector genome titer: gPCR; DNA quantitation; infectious particle titer)
  • Endotoxin testing (in development)


Lentivirus (HIV, SIV, and SHIV)
•    Virus stock production 

•    Virus titration (sMAGI; TCID50)

•    SIV plasma viral load (PVL) quantification (qPCR) 

•    Ultrasensitive SIV detection and quantification in tissues (nested digital qPCR)

•    SIV p27 protein and SIV antibody determination (ELISA)

•    SIV in vitro infectivity (leukocytes)

•    For lentiviral vectors, please contact disseng@ohsu.edu (Greg Dissen) at the MCB Core

Cytomegalovirus (CMV)
•    Virus co-culture from animal tissues

•    Virus stock and vector production 

•    Virus titration (TCID50)

•    Crude infected cell lysate for viral protein

•    Viral load quantification (qPCR) 

•    Antiviral antibody determination (ELISA)

•    Virus plaque purification

•    Virus growth curves

Expanded Simian-Pathogen Free Agents (ESPF)
•    RhCMV, RRV, and SFV viral antigen preparations for ESPF screening

Vaccinia (MVA)
•    Virus stock and vector production

•    Virus plaque assays

Miscellaneous
•    Small and large tissue processing and DNA/RNA extraction

•    Virus isolation from tissues and fluids

•    Manual and robotic nucleic acid isolation and purification

•    Cloning, recombination, and plasmid preparation

•    Western blotting to confirm viral protein or transgene expression  

•    Cell stocks, primary and cell lines

•    Sample processing and banking

The MVSC also offers data analysis and troubleshooting, custom services, consultation and training. Please inquire if you need additional assays not listed here and for assay development. We are committed to serving you and your research needs.  

Contacts


Christoph A. Kahl, Ph.D., Director, kahl@ohsu.edu, 503-200-4642
Don Siess, Ph.D., Senior Research Associate, siessd@ohsu.edu, 503-629-4042
Jeffrey Torgerson, Research Assistant 2, torgersj@ohsu.edu
Fax: 949-607-2608


Shipping Address


Molecular Virology Support Core (MVSC)
Research Building, Room 046
Oregon National Primate Research Center
Oregon Health & Science University
505 NW 185th Avenue
Beaverton, OR 97006


Facilities


The MVSC laboratories are located in the Research Building centrally located on the ONPRC campus. The MVSC laboratory (over 1,000 sq. ft.) contains a tissue culture room, as well as molecular virology and general laboratory space. The laboratory is divided into a Biological Safety Level (BSL)-2 space and a separate BSL-2+ laboratory (315 sq. ft.) used for work with infectious HIV and SIV samples. The laboratories are well equipped for virus isolation/propagation, PCR-based quantitative virology methods, serology/immunology, and molecular virology under BSL-2 and BSL-2+ containment levels. Major MVSC equipment includes biosafety cabinets, fume hood, CO2 incubators, fluorescent microscope, tissue grinders, robotic nucleic acid purification system, nanodrop spectrophotometer, table-top and ultracentrifuges, PCR hood, multifunction plate reader and washer, thermocyclers for quantitative and standard PCR, bacterial incubator and shaker, liquid nitrogen dewer, fridges and freezers for sample storage.