Duke University Regional Biocontainment Lab

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Contact Information

Director: Greg Sempowski, Ph.D
Email: greg.sempowski@duke.edu
Website: http://dhvi.duke.edu/diseases-research/shared-resources/regional-biocontainment-laboratory

Animal Capabilities, including species and containment level(s)

Enhanced ABSL3 housing available for mice, rats, guinea pigs, and rabbits.

Animal Models, including species, disease, and delivery method

All current models are in mice:

  • Yersinia pestis CO92, BSL3, aerosol, anterior cervical, footpad
  • Mycobacterium tuberculosis multiple strains (sensitive, MDR, XDR), BSL3, aerosol
  • West Nile virus IC7 & NY99, BSL3, footpad, intraperitoneal
  • Francisella tularensis SchuS4, BSL3, aerosol, intradermal
  • Vaccinia virus WR and Cowpox virus Brighton Red, BSL2, intranasal
  • Influenza virus PR8, BSL2, intranasal
  • Cryptococcus gattii and C. neoformans multiple, BSL2, aerosol

Pathogens

  • Current Select Agent approval: Bacillus anthracis Ames, Brucella abortis, Brucella suis, Brucella melitensis, Burkholderia pseudomallei, Francisella tularensis, highly pathogenic avian influenza, monkeypox virus, Yersinia pestis
  • Other agents handled in the facility: Mycobacterium tuberculosis (sensitive, MDR, XDR), Staphylococcus aureus, West Nile virus, vaccinia virus, cowpox virus, rabbitpox virus, human influenza viruses (H1N1, H3N2, B, H7N9), low pathogenic avian influenza virus, human rotavirus, vesicular stomatitis virus, yellow fever virus, Cryptococcus gattii, C. neoformans, N. gonorrhoeae, C. trachomatis

Imaging Capabilities

Spectrum In Vivo Imaging System (IVIS, Perkin Elmer) with both luminescence and fluorescence detection, validated for mice at enhanced ABSL3 (Appl Biosaf. 2010;15(2):62-66; PMCID: 3515055)

Laboratory and Analytical Support

All of the following are available at enhanced BSL3/ABSL3:

  • Immunology: In vitro functional assays, ELISA, live cytometry and cell sorting, multiplex biomarker analysis (Luminex), and real-time PCR arrays
  • Virology: Stock preparation and assay development (focus-forming units, TCID50, plaque-forming units); influenza hemagglutination inhibition and micro-neutralization assays
  • Bacteriology: Inoculum preparation and validation, colony-forming units
  • Pathology services: Sample collection and necropsy for tissue collection/fixation, access to Duke Department of Pathology core lab for embedding/cutting

Ability To Accommodate cGLP or cGMP Studies?

  • Current GLP: HIV propagation and assays with quality control for proficiency panels; Luminex biomarker assays and Luminex proficiency testing (accredited to the ISO/IEC 17043:2010 standards by the American Association for Laboratory Accreditation [A2LA])
  • Current cGMP: None

Specialized areas or Major Equipment Available

All of the following specialized resources and major equipment are installed, validated, and available at enhanced BSL3/ABSL3:

  • BD FACSAria IIu cell sorter; Luminex bead array reader; real-time Thermal Cycler, ELISA plate reader and washer
  • Two whole-body aerosol exposure systems, each with capacity for simultaneous exposure of 144 mice, 18 guinea pigs, 6 rabbits, or similar numbers of other small animals and one nose-only exposure system for exposure of 36 mice.
  • Aerosol Management Platform (AeroMP, Biaera) to control whole-body and nose-only exposures (controls and records flow, pressure, humidity, and temperature)
  • Aerodynamic Particle Sizer (APS, TSI) for real-time monitoring of particle number and size distribution (count or mass) in infectious aerosols.
  • Biosamplers and impingers to determine concentration of viable bacteria or yeast in infectious aerosols.

Shared Resources That Amplify and Expand Research Capabilities

DHVI Shared Resources/Core Labs (Scientific Director: Gregory D. Sempowski, Ph.D.): The faculty and senior scientific staff of the Duke Human Vaccine Institute have assembled state-of-the-art technologies into Shared Resources (i.e., Core Labs) to accelerate research. These comprehensive Shared Resources are available to the entire Duke community and their collaborators. DHVI cores are supported with institutional funds and a variety of shared instrumentation grants, center grants, and program awards.

The RBL at Duke functions administratively as one of the eight DHVI Shared Resources and this membership allows RBL users access to a wide-range of cutting edge technologies and facilities to enhance their research.

  • Regional Biocontainment Laboratory: Home to a portfolio of collaborative research units focusing on Bacteriology, Virology, and Immunology. Fully-commissioned for BSL3/Select Agent work in late 2007 and has supported morethan 60 publications.
  • Research Flow Cytometry & Cell Sorting: Analytical/sorting on six BD instruments with up to 23 simultaneous parameters.
  • Biomolecular Interaction Analysis: State-of-the-art Surface Plasmon Resonance (SPR) BIAcore instruments for monitoring real-time interaction analyses and measurement of binding affinity, kinetics, and epitope mapping.
  • Immunology Virology Quality Assessment Center Laboratory: Delivery of CAP- and CLiA-certified immunologic and viral load assays (i.e., immunophenotyping; SIV viral load, HIV serology, HIV and HCV viral load; HIV acute infection disease staging, and large-scale cell separation.
  • Viral Genetic Analysis: Large-scale DNA sequencing, Next-Gen sequencing, and sequence data analysis, to serve the viral genetic analysis needs of researchers.
  • X-ray Crystallography: Automated crystallization systems and high resolution X-ray diffraction image collection to solve macromolecular crystal structures (crystallization trials, data collection, phasing, refinement, and analysis).
  • Protein Production Facility: Quality and standard recombinant protein production under “GxP best practices” for in vitro use.
  • Accessioning Unit and Clinical Trials Support: Centralized support for preparation of human clinical studies, collection of human specimens, and ongoing compliance oversight. Processing and continuous auditable storage/monitoring of frozen samples derived from studies/trials.

Website for Shared Resources: http://shared-resources.dhvi.duke.edu/​

Content last reviewed on November 14, 2016