Biological Safety FAQs

Recombinant DNA (rDNA) was originally created by cutting DNA segments with restriction enzymes, then re-combining them in a ligation reaction. Today, rDNA is commonly generated by PCR, de novo synthesis, and other methods. The NIH guidelines cover both rDNA and RNA that is derived from rDNA. The Duke IBC includes the following as rDNA materials: 

• Plasmids and viral vectors 
• Any synthetic DNA or RNA 
• Any RNA produced from rDNA, including messenger RNA (mRNA), small interfering RNA (siRNA), micro RNA (miRNA), etc. 
• Genetically-modified organisms (animals, plants, bacteria, viruses, fungi, etc.) 

The Duke IBC defines a viral vector as any rDNA molecule or molecules used to deliver nucleic acids to cells using viral proteins or their equivalents for cell entry. Examples of viral vectors include replication-deficient forms of adenoviruses and retroviruses. Typically one or more plasmids are transfected into cells. The plasmids express viral proteins in the cells. These cells produce replication-deficient viral particles containing both nucleic acids and viral proteins. The replication-deficient viral particles are used to efficiently deliver the nucleic acids to other cells either in vitro or in vivo. Replication-competent viruses are sometimes used as hosts for recombinant DNA research.   

A genetically-modified organism (GMO) is any organism whose genetic material has been altered using rDNA technology. Note that descendants of GMOs are also considered GMOs. The term GMO includes any form of life (vertebrate or invertebrate animal, plant, bacteria, fungus, virus, etc.). Common examples of GMOs include transgenic mice, knockout mice, and bacteria used to make recombinant proteins. 

All research at Duke University and Health System using recombinant DNA (rDNA) materials must comply with the NIH Guidelines for rDNA Research. Each investigator working with rDNA is responsible for understanding and following the NIH guidelines. Take the rDNA survey to determine if your work requires IBC registration. If determined non-exempt, then the work must be registered with the IBC. This includes work with rDNA materials obtained from other scientists or from commercial sources. 

Experiments involving the deliberate formation of recombinant or synthetic nucleic acid molecules containing genes for the biosynthesis of toxin molecules must be registered.  

For non-clinical studies, complete the IBC Recombinant DNA Registration form and submit it via e-mail to the Duke Biological Safety Office (biosafety@dm.duke.edu). Other information, such as a written SOP, may be required depending on the research involved and biocontainment level. Refer to documents under Quick Links on this page. 

For clinical studies, navigate to the Duke iRIS system to register. Ideally, the IBC registration form is submitted simultaneous in the IRB submission. However, you may submit the IBC registration form separately in iRIS. An Investigational Product Handling Plan (IPHP) may be required with the submission.  

Each rDNA registration requires a descriptive title, and should cover a single area of research. Researchers who conduct rDNA research in multiple areas should have multiple registrations.  

Each rDNA registration has a single PI who takes responsibility for assuring compliance for all work on the registration, and for assuring that all workers in all locations are appropriately trained to conduct the research. 

Each rDNA registration must list every location where rDNA will be manipulated under the registration. Existing protocols can be amended to add new locations. The procedures for safe work with rDNA, as described in the registration, must be followed by everyone listed on the protocol in all locations.  

For non-clinical studies, use your existing IBC registration and make modifications or additions to the work, while highlighting the newly added text. Any new viral vectors or other biological materials for work at BSL-2 will need to be added to your SOP, while highlighting the newly added text. Submit documents to biosafety@dm.duke.edu. 

For clinical studies, navigate to the Duke iRIS system to submit the IBC amendment form.  

Registrations require an annual review and renewal date. PIs will be notified annually and asked to complete an annual review form for continuing projects to document that information regarding non-exempt investigational gene therapy or vaccine products, gene constructs, work procedures, animal use, or other significant changes are current with the IBC records. 

A single registration can cover collaborative research in more than one lab if ALL the following conditions are met. (1) The collaborative research is described in the registration. (2) All locations and faculty members are listed on the registration. (3) All work is conducted in compliance with the IBC registration including any Standard Operating Procedures (SOPs). (4) The PI of the registration assumes responsibility for assuring that all work in all labs is compliant with the registration, and for assuring that all workers in all locations are appropriately trained to conduct the research. Existing protocols can be amended to describe collaborative research and to add additional faculty members. 

Any potential exposures will be reported immediately by calling the Employee Occupational Health and Wellness (EOHW) exposure hotline (919-684-8115). EOHW will evaluate each exposure and develop an appropriate response. Any accidental exposure to the rDNA material will be reported to the OESO-Biological Safety Division (919-684-8822) who will report to NIH according to the NIH rDNA Guidelines. 

• Eye exposure from splash or aerosol 

Rinse a minimum of 15 minutes in eye wash or flush with water. Immediately notify EOHW by dialing 115 from a Duke Phone or 919-684-8115 from any phone.  

• Needlestick, sharps exposure, or non-intact skin exposure 

Contaminated skin must be washed with soap and copious amounts of water. Immediately notify EOHW by dialing 115 from a Duke Phone or 919-684-8115 from any phone. 

When a patient is on Airborne Infection Isolation (AII). In order to enter the AII room, you need to put on respiratory protection: either a N95 or PAPR. For more information on these, please see the TB ECP.  

A patient is put on Airborne Infection Isolation when they have a suspected airborne illness. The patient is put into an Airborne Infection Isolation (AII) room. An AII room is a negative pressure room: this means that air flows from the hallway into the anteroom, and from the anteroom into the patient room. The air cannot flow outward from the patient room to the hallway, or negative pressure would be lost. Therefore, the door needs to be closed at all times when a patient is on AII.  

A tissue test is a method to test the negative pressure of the AII room. Take a small piece of tissue and hold the tissue approximately 1 inch above the floor at the bottom of the closed door. If the room has a sliding glass door, you can slightly crack (<2 inches) the sliding door in order to perform the test. If the air pressure is sufficiently negative, the tissue will be sucked toward the room. A tissue test needs to be done every day that a patient is on AII in the room.  

If an AII room is not available, call for a Portable HEPA unit (see Appendix D in the TB ECP). After the patient is moved to an AII room, follow the guidelines below:  

1. If a patient wore a surgical mask at all times, then routine procedures can be used to clean the room.  
2. If a patient did not wear a surgical mask and a HEPA unit was used, the door must be closed and the room empty for 30 minutes.  
3. If a patient did not wear a surgical mask and a HEPA unit was not used, the door must be closed and the room empty for 120 minutes.  

Notify Biological Safety (biosafety@duke.edu) or Employee Health (919-684-3136).  

Once a patient is confirmed to have TB, we will follow up with Tuberculin Skin Testing, as needed. For complete post-exposure procedures, please see the TB ECP.   

Please reach out to biosafety@duke.edu.   

Depending on your location, see our ECP for Hospital and Clinics, Clinical Research Laboratories, Private Diagnostic Clinics, and Duke Primary Care or our ECP for Research Laboratories and Other Non-Healthcare Areas. In addition, review our training requirements. 

For work with human blood, body fluids, and other potentially infectious material (OPIM), such as human cell lines see our Universal/Standard Precautions for Handling Human Specimens. An additional risk assessment is required for known or suspected prion disease tissue. For questions, contact OESO – Biological Safety Division at (919) 684-8822 or via biosafety@duke.edu.  

See our Biological Exposure and Spills webpage for detailed instructions.  

Regulated medical waste means blood and body fluids in individual containers in volumes greater than 20 milliliters, microbiological waste, and pathological waste that have not been treated. 

Biological waste includes untreated regulated medical waste and biological materials generated from research, diagnostic, and/or teaching activities during manipulation or cleanup procedures.  

No. OESO-Environmental Programs collects chemical, pharmaceutical, and radiological waste.  

Review the information within the Duke Medical Waste Management Policy for guidance specific to the types of waste and your work location.  

• In the hospital or medical center buildings: Environmental Services (EVS) at 919-681-9700.  
• In other locations, contact your departmental business office for information on setting up a Stericycle pickup service. Review the information within the Duke Medical Waste Management Policy for guidance specific to the types of waste and your work location. 

Put on PPE to cover all skin, eyes, nose, and mouth. Gather the components of your spill kit. Follow the instructions in the BIOLOGICAL section of the Emergency Response and Incident Reporting Guide. 

• Eye/face exposure
  Rinse a minimum of 15 minutes in eye wash or flush with water. Immediately notify EOHW by dialing 115 from a Duke Phone or 919-684-8115 from any phone.  

• Needle stick, other sharps exposure, or non-intact skin exposure
  Contaminated skin must be washed with soap and copious amounts of water. Immediately notify EOHW by dialing 115 from a Duke Phone or 919-684-8115 from any phone. 

• Any machine that processes blood, body fluids, or other potentially infectious materials (OPIM).
  • OPIM includes semen, vaginal secretions, cerebrospinal fluid, synovial fluid, pleural fluid, pericardial fluid, peritoneal fluid, amniotic fluid, saliva, any body fluid contaminated with blood, any unknown body fluid, or any unfixed tissue or organ except intact skin.
• Containers of regulated medical waste.
• Freezers, incubators, or holding areas for blood, body fluids, or other potentially infectious material (OPIM).
• Other containers or bags used to store or transport contaminated equipment, materials, needles, and sharps, such as:
  • Sharps containers (if not already labeled).
  • Biohazardous waste collection bins.

• Containers must be wall mounted or placed on a stabilizing tray.  
• Containers must be upright and easy to use while preventing the contents from spilling.  
• Containers must be placed in a visible location, within easy horizontal reach, and below eye level (according to NIOSH standards).  
• Containers must be placed away from any obstructed areas, such as near doors, under sinks, near light switches, etc. (CDC).  

No, in this case, the red color of the bins is sufficient to indicate a biohazard container. 

No, assigned blue bag is sufficient to identify soiled linen.  

• No, red bags are not required.  
• Soiled linen should be placed in a fluid resistant bag. Blue bags are acceptable.  
• Linens soaked with fluids should be double-bagged.  

• Depends on the amount stored in the room or the location. Joint Commission requirements state: 
  • Rooms that store more than 64 gallons of trash or soiled linen have doors that are self-closing and latching (LS.02.01.30 EP 2). 
  • Soiled utility rooms in behavioral health units are required to be self-closing and self-locking (NPSG.15.01.01 EP 1). 
• If neither of the above conditions apply, soiled utility rooms do not need to be locked. 

Yes, contact OESO at biosafety@duke.edu 

No, for use in patient care or clinical care areas, all biohazard warning signs and labels for use in these areas must be distributed by OESO.  

For laboratories, you may order the labels so long as they meet the OSHA BBP Standard requirements:  

Labels must be fluorescent orange or orange-red or predominantly so with the biohazard symbol and the word “BIOHAZARD” in a contrasting color. 

Contact OESO at biosafety@duke.edu 

In laboratories, biohazard labels are required on sharps containers that contain infectious substances or human material. 

In patient care or clinical care areas, if the sharps container is not red, a biohazard label is required. 

• Red is preferred.  
• It can be any color but do not use blue containers to avoid confusion with the blue bin program (pharmaceutical waste) containers.  
• If it is not red, a biohazard label is required.  

Anyone who classifies materials, prepares paperwork, and/or prepares packages of biological materials for shipping must complete appropriate training at least every two years. 

The U.S. Department of Transportation Hazardous Materials Regulations (HMR) have aligned with international regulations (ICAO/IATA). If you follow the IATA DGR, you will be in compliance with both international and domestic requirements. 

You need to take OESO Shipping Biological Materials Training. Exempt patient specimens (containing no other hazardous materials) for which there is minimal likelihood that pathogens are present are not subject to other shipping regulations, but must be packaged as Exempt Patient (human or animal) Specimen using the checklist in the Training Supplement Guide. 

Biohazard and potentially infectious material sample transportation is PROHIBITED on public                      transit, but vehicles dedicated for patient specimen transport (Duke Clinical Labs) where no passengers are inside the vehicle may be used.   

Specimens may be transported by private or contract carrier vehicle that used exclusively for these materials. When lab specimens are transported across a public street or highway they are covered by the DOT requirements and must be packaged accordingly. Transporters must have shipping and spill response training. Your department is responsible for providing approval. OESO does not approve the use of private vehicles for transporting infectious material. 

An infectious substance which does not meet the criteria for inclusion in Category A. Infectious substances in Category B must be assigned to UN 3373. 

An infectious substance which is transported in a form that, when exposure to it occurs, is capable of causing permanent disability, life-threatening or fatal disease in otherwise healthy humans or animals. Indicative examples of substances that meet these criteria are below.