McMaster University

McMaster University

UpdatedBiosafety Level 2 Plus Lab Preparation and Training

Description and Prescription of CL2+ Containment

Biosafety Level "2+" or "2 enhanced" is the name is given to the scenario whereby a pathogen or sample is deemed to require CL2 physical containment using additional operational procedures beyond those prescribed for CL2. Basic CL2 operational procedures are outlined in the Canadian Biosafety Standard. The pathogen is handled in a BSL2 physical space with BSL2 equipment.

The decision for a given pathogen to be handled at CL2+ will come from one of three sources:

(1) The import permit under which the pathogen was imported requires it (see restrictions on your permit)

(2) A formal Risk Assessment by the Presidential Biosafety Advisory committee has found that these precautions are warranted

(3) The above Risk Assessment was inconclusive and a query was forwarded directly to the Office of Laboratory Biosafety, PHAC and they have deemed that these precautions are warranted

CL2+ is required for (but not limited to):

  • Diagnostic samples and cultures of HIV
  • Diagnostic samples and cultures of HTLV
  • Diagnostic samples of Hepatitis C Virus
  • Diagnostic samples of West Nile Virus
  • Diagnostic samples of Mycobacterium tuberculosis
  • Samples derived from primates within the genus Macaca
  • Diagnostic samples of SARS-CoV-2 (Covid19)
  • Lentiviral vectors rated at BSL2+ per Risk Assessment PID


The additional operational procedures specific for the above samples and a more detailed explanation is found in the new PBAC Implementation Directive - Operational Procedures for Use at BSL2+.


Getting Your BSL2 Laboratory Ready for BSL2+ Work

Physical Space

Your laboratory must be suitable for BLS2 work. In practice these features are iterated by the Canadian Biosafety Standard.

The items appropriate for a BSL2 laboratory are found in the document used for the audit of your BSL2 laboratory space, the Combined Checklist. To ensure that your laboratory complies with this checklist, please contact the Research Compliance Auditor (link on left) for help and suggestions.


Working under BSL2+ operational protocols requires a complete set of SOPs to be written. The SOPs required include but are not limited to:

(1) Layout of the BSL2+ area when in BSL2+ mode, Laboratory Access, Biosecurity

(2) PPE Required, Entry and Exit Protocols, Decontamination, Housekeeping

(3) Emergency Exit Protocols

(4) Spill Protocols

(5) Lab Specific protocols (experimental procedures done at BSL2+)

(6) Use of a Biological Safety Cabinet

At any time during this process, you may contact the Research Compliance Auditor (link on left) in the Biosafety Office for SOP review and comments. When completed, the set of SOPs are submitted to the Presidential Biosafety Advisory Committee for review.

Solid Front, Rear Tying, Tight Wristed Gowns

If required, options for these types of gowns are disposable or rental. For rental gowns, you must create an "on demand" account at Mohawk Linen Services by contacting customer service representative Randy Spencer. The account allows you to receive clean, non sterile gowns in bulk which you then send back in bulk for laundering. The type of gowns to order are called "Polygown Class B" (one size fits all) and you will require a "nylon bag" for return of dirty gowns. Gowns from Mohawk are delivered to the MUMC Receiving docks in the Health Sciences Centre. Contact Giulio Paiano, Hamilton Health Sciences Customer Support Services Leader (905-521-2100 x76440), for manual drop off and pickup locations or to create an account for trucking options.

If a gown or other rental linen becomes contaminated due to a spill, please bag the gown securely in an autoclave bag and contact the Biosafety Office. Do not bleach. Do not bag with uncontaminated gowns.


Training for BSL2+ Work

    Site and work-specific training are required by the Canadian Biosafety Standards and Guidelines. Every person (workers and Supervisors) must have completed BSL2 training, WHMIS and Fire Safety and all the associated updates. There are four additional tasks that need to be completed:

    (1) The new worker must be familiar with the physical space that is to be designated the BSL2+ work area. The Supervisor should assure this.

    (2) The Supervisor must ensure the new worker is trained on the SOPs required for their BSL2+ work:

  • general SOPs
  • project-specific SOPs
  • spill cleanup SOPs


This documentation will be audited annually.

    (3) The new worker should understand the nature of the pathogen with which they are working. This information should be provided by the Supervisor. The Pathogen Safety Data Sheets may be found here.

    (4) N95 respirators are required to be available in the event of a spill. Contact the FHS Safety Office (x24956) or EOHSS (x24352) to arrange for respirator training and N95 mask fit testing. Fit testing training information can be found here. The Supervisor is also expected to complete N95 training and fit testing since the expectation is that the Supervisor should be available to directly help or directly supervise the worker during a spill cleanup. If the Supervisor physically cannot directly help in spill cleanup, the medical accommodation procedure must be followed. If you have an accommodation need, please email or call x21694 to make appropriate arrangements.

    When the worker and Supervisor have completed all training listed above, they may be deemed competent to work safely at BSL2+.

    Regular Lab Audit

    All biohazard labs are audited regularly according to the schedule found here. A BSL2+ lab is audited to the CBS containment level 2 standards. Additionally, the Research Compliance Auditor will look for the following:

  • review the information provided on the BUP
  • review the training records of each person listed on the BUP
  • inspect BSL2+ SOPs to ensure they are reviewed annually. This can be indicated by a date/signature reflecting the last review. There is no need to reprint the SOPs annually if there are no changes.
  • inspect training documentation for all BSL2+ workers. The expectation is that each worker has been trained on the the BSL2+ SOPs. The Supervisor is responsible to create and deliver the training to the workers. Documentation of this training will be audited. Documentation may take any form as long as it contains a signature from both the worker, the Supervisor and the date on which the training took place.
  • inspect the spill kit
  • verify appropriate N95 respirators are available in the working areas


    Updated: 24-Mar-2020

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