- BSL-3 Facility
- Training Program in HIV Biosafety Practices for Lab Personnel:
- Part 1: Manual, General Trainings, and BSL-3 Video
- Part 2: Hands-on Training Program (60-minute session)
Training Part 2: Hands-on Training Program (60-minute session)
1. Laboratory Entry/Exit procedures: Gowning, Double gloves, Arm Covers, clothing, shoes
Goal: This session reviews the principles of barrier maintenance between the user and work environment by gowning, double gloving and arm covers. The double gloving is shown directly, with users exchanging the first and second pair of gloves for laboratory entry and work environment usage. Proper glove using is reviewed, as well as common misconceptions about glove barriers. Users also learn what impact rinsing gloves with 70% alcohol or detergent has on rubber integrity. Guiding principles on reusing gowns, and what represents a soiled gown are also addressed.
Activity: Undertake a complete gowning, double gloving and arm covers. Time: 5 minutes.
2. Biosafety Cabinet Setup and Waste Management
Goal: This session reviews set up of a biosafety cabinet for work with HIV infectious material. Management of (a) liquid waste, (b) open waste such as pipettes and (c) sealable waste such as tissue culture flasks are covered, along with proper setup of disposal.
Activity: Set up the hood with proper waste management options while reviewing understanding of the principles behind each waste category. Time: 15 minutes.
3. Concepts of Work Areas, Glove Changing
Goal: This session reviews the identification of different work areas within a BSL-3 environment. The immediate area of the hoods has the highest potential for exposure while other areas such as telephone, note taking and computer areas can be considered secondary work areas via change of double gloves between areas. Entry into the BSL-3 when not working in hood area is also discussed if this work-area strategy is used.
Activity: Users are shown representative work areas within the environment and the proper way to transit between them, as well as how to answer the phone and use a computer when coming from a biosafety hood area. Time: 5 minutes.
4. Centrifuge Principles
Goal: This session reviews adequate sealing and transport between the hood and centrifuge, and potentials for exposure when spinning micro tubes versus whole size tubes. Use of micro-centrifuges and regular area centrifuges is covered. A spinning session will be demonstrated, following the splitting of non-adherent cell lines using the waste setup reviewed in a prior session. This will also allow to apply operational waste management handling as reviewed in a previous session.
Activity: Hood cell splitting and centrifuge usage in a BSL-3 environment. Time: 15 minutes.
5. Ending Work Session and HIV Deactivation
Goal: This session reviews the principles of chemical or heat inactivation of HIV-1 particles. Waste management is addressed, including deactivating solution and waste removed from the biosafety hood. Expectations for autoclaving and waste management for the environment are also covered. Oven system deactivation is covered including deactivation via detergent and heat by 56 degrees over 20 minutes or 95 degrees within a two-minutes PCR cycle.
Activity: Cleaning the hood area with Virkon or Clorox dilution, removing waste from the biosafety cabinet and preparing hood work station for the next user. Oven heat deactivation system is shown. Time: 10 minutes.
6. Transport Principles
Goal: This session reviews principles for transporting infectious material outside of the BSL-3 environment. A two-container system is shown along with a demonstration of the approach to using this container.
Activity: Packaging of infectious material for transport outside of the BSL-3. Time: 5 minutes.
7. Accident and Emergency Procedures
Goal: This session reviews actions to be taken in case of an accident or emergency. The user learns the location of the occupational health office and what are the current approaches to post-exposure prophylaxis. Users are made aware of the need for emergency contact numbers to be accessible as well as laboratory safety manual (users will be shown these). In addition, we review actions if a spillage occurs outside a biosafety cabinet, within the microscope, centrifuge or bench area. Users are also alerted as to how radioactivity may impact emergency procedures.
Activity: Review of the location of the Occupational Health Office and how this office can be reached by telephone or by foot ( Contact information here ). Review of emergency number listings in the laboratory and access to emergency procedures hand-book. Time: 5 minutes.