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ABSL-3/BSL-3 Biocontainment Core

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Research on microbial agents, which cause lethal diseases in humans and for which effective drugs or preventive vaccines are not available, must be conducted by well-trained investigators in specially built, well-maintained laboratories. The BSL-3/ABSL-3 Biocontainment Core provides the triad of service, research and development, and education and training to investigators at the university and the wider research community.

Objective

Enhance the infrastructure for the BSL-3/ABSL-3 Biocontainment Core, in support of hypothesis-driven research projects that seek to develop rapid diagnostics, more effective treatments and affordable preventive vaccines for diseases caused by BSL-3 agents.

Specific Aim 1

  • Enhance support and services in BSL-3/ABSL-3 biocontainment.

Specific Aim 2

  • Develop innovative and improved microbiological assays for research on BSL-3/ABSL-3 microbial pathogens.

Specific Aim 3

  • Develop and implement training and education workshops in biocontainment-related technology.
People
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Vivek R. Nerurkar, Ph.D.

Director, Biocontainment Core
Professor and Chair
Department of Tropical Medicine, Medical Microbiology and Pharmacology
John A. Burns School of Medicine


Email: nerurkar [at] pbrc.hawaii.edu
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Mukesh Kumar, M.S., Ph.D.

Associate Director, Biocontainment Core
Instructor
Department of Tropical Medicine, Medical Microbiology and Pharmacology
John A. Burns School of Medicine


Email: mukesh [at] hawaii.edu

Services/Assays

BSL-3 assays and services

  • West Nile Virus plaque assay: Titration of serum samples or tissue homogenates on Vero cells, using traditional agar overlays and neutral red. Calculate virus titers using Excel or your specified output file.
  • Homegenate preparation for plaque assay or ELISA: Homogenization of tissue samples using the bead-based Bullet Blender or Fastprep. Clarified samples stored frozen in the facility until used for the plaque assay, ELISA, or other assays.
  • West Nile virus PRNT assay: Determination of PRNT90 values for serum samples. Raw data and calculated values returned to the user in Excel or specified output file.
  • Luminex assay: Plate set-up, incubation of your samples with beads from your kit, and running of plate on the Luminex 200 instrument in our facility. Raw data files returned in Excel or your specified output file. Samples accepted for testing include: serum, 0.4 microM filtered tissue homogenate, and cell supernatant. Other specimens subject to approval from the Laboratory Director.
  • Virus production: Culture, titration, and working dilution determination of West Nile virus or other IBC approved virus. Vials will be stored in our facility until needed for assays.
  • Nucleic acid isolation: Isolation of total RNA or genomic DNA for downstream analysis such as RT-PCR. Concentration of each sample determined using UV spectrometry.
  • RT-PCR: Plate set up, cDNA synthesis, and running of samples with primers and probes on the real-time PCR instruments. Five different color detection options. Quantitative or relative analysis available.
  • Flow cytometry: Creation of single cells suspensions from your tissue samples using the Gentle MACS system, and staining of cells with antibodies. Samples will be fixed and run on the FACS Aria flow cytometer. FACS Aria charges are billed separately.
  • Freezer rental space: Affordable monthly rentals space on a per box basis.
  • Other services: Cell migration assays, cryostat sectioning, fluorescent microscopy, and protein isolation. Please contact us for more information, or if you may require an assay or procedure that are not included in this list. Subject to authorization by the Biocontainment Laboratory Director.

ABSL-3 assays and services

  • 21-day survival study: Inoculation of mice with West Nile virus and monitoring of symptoms for three weeks. Includes chain of custody, husbandry, removal of carcasses, and terminal bleed out of survivors.
  • Eight-day comparison study: Inoculation of mice with West Nile Virus and monitoring of symptoms for eight days. Includes chain of custody and husbandry.
  • Tissue harvest with perfusion and/or paraformaldehyde tissue preservation: Perfusion and collection of blood/serum and specified tissues. Tissues are flash frozen in liquid nitrogen or fixed in formaldehyde and embedded in OCT medium for sectioning. Rates charged are per harvest.
  • IP injection: Animals will be administered doses of your IBC/IACUC approved compound via intraperitoneal injection route. Rates charged are per injection.
  • Blood collection via tail-bleed: Less than 100 uL of blood will be collected via tail-bleed from mice according to the approved IACUC protocols, and then centrifuged to freeze serum for downstream analysis. Rates charged are per collection.
  • Tissue harvest (non-perfused): Collection of blood/serum and specified tissues. Tissues are flash frozen in liquid nitrogen. Rates charged are per harvest.
  • Evans Blue Assay: Injection of mice with Evans blue dye for blood-brain barrier permeability assessment. Services include images and scoring of each sample.
  • Other services: Intracranial injections and survival surgeries. Please contact us for any services you may require that are not included in this list. Subject to IBC/IACUC approval and authorization by the Biocontainment Laboratory Director.

Training opportunities

Experienced staff will provide hands-on training to investigators in conducting IBC/IACUC approved BSL-3/ABSL-3 research protocols. Please refer to our training document (Training Required for New Personnel) to help you get started on the process for obtaining clearance to work in the JABSOM Biocontainment Facility.

Equipment

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Protocols/Procedures

Please fill out the service request form and email it to the ABSL/BSL-3 Biocontainment Core.

Publications

Citations in Publications

Use of this core facility should be acknowledged in publications, abstracts, posters and oral presentations. The suggested verbiage is:

“Some of the services for this research were provided by the ABSL-3/BSL-3 Biocontainment Core, which is supported in part by grant P20GM103516 from the Centers of Biomedical Research Excellence (COBRE) program of the National Institute of General Medical Sciences, a component of the National Institutes of Health.”

2011-2012

2010-2011

2009-2010

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