Pathogens Unit at the RBL

As a core service laboratory, the RBL Pathogens Unit of the Regional Biocontainment Laboratory at Duke University is tasked with assay development, optimization, and execution in order to provide high quality, well-controlled and reproducible research that is delivered in a professional and efficient manner. We also provide training, reagents, protocols, and consultation to investigators.

The Pathogens Unit has a robust and active influenza virus support program as well as a significant SARS-CoV-2 support infrastructure. We provide virus stock propagation and quality control as a service, and deliver that material to researchers for use in animal models, in vitro testing, and as standards for assay development. Additionally, the Pathogens Unit possesses the expertise and experience to quickly onboard new pathogens of interest.

The core modality of the group is to assess the neutralization of live virus (Microneutralization Assay, Hemagglutination Inhibition Assay, or Plaque Reduction Neutralization Test) in response to various immune sera or monoclonal antibodies submitted by intra- and intermural investigators, at both BSL2 and BSL3. The Pathogens Unit routinely measures viral load in a variety of bio-specimens (Plaque Assay, Tissue Culture Infectious Dose, or Quantitative Real Time PCR). Our influenza inventory contains over 80 strains isolated in the past 100 years. We continue to acquire and propagate SARS-CoV-2 variants of concern as each arises.

 

RBL Virology Unit, from left to right: Kamerin Harvey, Madeline Hayes, Michelle Rock, MS, Trey Oguin, PhD, Emily Hill and Jessica Huskey
RBL Pathogens Unit, from left to right: Kamerin Harvey, Madeline Hayes, Michelle Rock, MS, Trey Oguin, PhD, Emily Hill and Jessica Huskey
Director of RBL Pathogens Unit:  

Brook Heaton, PhD

(919) 681-3299

Unit Manager:  

Trey Oguin, PhD

(919) 681-3072

Quality-controlled virus stock propagation

  • BSL2, BSL3 and Select Agent
  • Tissue culture propagation

Viral load quantification

  • Plaque/focus forming unit assays
  • Tissue culture infectious dose 50% assays
  • Hemagglutination units assay (Influenza)
  • Subgenomic qRT-PCR for SARS-CoV-2

Virus serology assays

  • Hemagglutination inhibition assay (Influenza)
  • MicroNeutralization assay
  • Plaque reduction neutralization test

In vivo challenge models including sample procurement and processing from infected animals (see Animal Models Unit)

  • Human influenza in ferrets
  • Mouse-adapted influenza in mice
  • Rabbitpox in rabbits
  • SARS-Cov-2 (in development)

Development of project-specific virology support assays

  • Track record working with flaviviruses, human cytomegalovirus, influenza viruses, coronaviruses, human rotavirus, poxviruses, and vesicular stomatitis virus

Small molecule inhibitor characterization- BSL2/3

  • Minimum Inhibitory Concentration assay

  • Small molecule inhibitors and compound testing (EC50 and IC50 determination) 

Bacterial Culture

  • Colony forming unit assay

  • Quality controlled bacterial stocks propagation: BSL2, BSL3, and select agents

  • Mycobacteria isolation from human tuberculosis specimens (sputum), stock propagation, genomic DNA, and RNA isolation

Cell culture work for BSL2/BSL3-specific agents

  • Isolation of Bone Marrow-Derived macrophages

  • Differentiation and activation of mammalian cells (e.g., THP1 cells)

  • Infection with mycobacteria

  •  RNAi-based experiments in cells

Endotoxin testing

  • Endotoxin testing

Microplate Assay

  • Microplate reader-based bacterial quantification controlled with Gen5 data analysis software interface.

  • Gene amplification and qRT-PCR assay (96 well and 384 wells)

Bacterial and fungal challenge model in mice

  • The Pathogens core has established bacterial doses for infecting animals with Mycobacterium tuberculosis, Yersinia pestis, and sexually transmitted diseases e.g., Chlamydia spp., and fungal agents e.g., Coccidioides immitis, etc. We provide customized full or partial support to meet your microbiology lab and animal model needs, including aerobiology (see Animal Models). 

Assay Request & Sample Submission Tool

Step 1:  Download and complete the appropriate sample submission manifest
Step 2:  Complete the Assay Request Form 

Use Fees/Rates

Current approved rates for Duke Investigators can be seen in CoreResearch@Duke (Core D-0031).

CoreResearch@Duke can also be used to request an estimate for a grant or contract proposal.

Service Requests and Estimates

External Non-Profit Institution users:

  • A signed Research Analysis Quote and Agreement (RAQA) and Purchase Order (PO) will be needed from your institution.
  • Contact the Pathogens Unit for a price quote to get the process started.

Non-Duke For-Profit Entity users:

  • Duke Shared Resources can only provide services to For-Profit Entities under a signed Sponsored Research Agreement (SRA)
  • The collaborative research project must have a defined research question, scope of work, budget, and timeline.
  • Contact the Pathogens Unit to get the process started.

Influenza Assay / Animal Model Request (form link)

  • To be used by CIVICs investigators to submit requests for animal studies and RUO assay support
  • Download required RUO assay sample submission manifest 

CIVICs Assay Guidance

Please Note:   These links/forms are for the exclusive use of Duke CIVICS program investigators.

Directions to the RBL and Parking Information (contact us)

How to acknowledge work done in our facility:

Please acknowledge our facility in any publications/presentations:  “____ was provided under the management of Dr. Thomas "Trey" Oguin, III and direction of Dr. Brook Heaton in the Pathogens Unit of the Duke Regional Biocontainment Laboratory (RBL), which received partial support for construction from the National Institutes of Health, National Institute of Allergy and Infectious Diseases (UC6-AI058607; G20-AI167200) and facility support from the National Institutes of Health (UC7-AI180254).”