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The Laboratory Biosafety Guidelines: 3rd Edition 2004

Chapter 4
 Laboratory Design and Physical Requirements

This chapter is designed to provide guidance on the design and layout required to achieve the four containment levels detailed in Chapter 2.

The chapter is divided into five matrices: Laboratory Location and Access; Surface (i.e., floors, walls, ceilings, sealants) Finishes and Casework; Heating, Ventilation and Air Conditioning (HVAC); Containment Perimeter; and Laboratory Services (i.e., water, drains, gas, electricity and safety equipment). Information on commissioning, certification and recertification of the containment features detailed in the matrices can be found in Chapter 5.

Matrix Legend: Mandatory - mandatory Recommended - recommended

4.1 Matrix 1
Laboratory Location and Access

Matrix 1 Containment Level Laboratory Location and Access
1 2 3 4
1 Mandatory Mandatory Mandatory Mandatory Separated from public areas by door.
2   Mandatory Mandatory Mandatory Access limited to authorized personnel.
3   Mandatory Mandatory Mandatory Laboratory room doors to have appropriate signage (e.g., biohazard sign, containment level, contact information, entry requirements).
4 Mandatory Mandatory Mandatory Mandatory Size of door openings to allow passage of all anticipated equipment.
5   Mandatory Mandatory Mandatory Doors to the containment laboratory lockable (this does not apply to areas within the containment laboratory).
6     Mandatory Mandatory Doors to provide restricted access by installation of a controlled access system (e.g., card key) or equivalent.
7     Recommended Mandatory Electronic locking systems to be backed up with a physical key-lock system.
8   Recommended Mandatory Mandatory Office areas to be located outside of containment laboratory. Paperwork stations for data collection can be within containment laboratory provided they are located away from laboratory work areas.
9     Mandatory Mandatory Entry to laboratory to be provided via an anteroom.
10     Mandatory   Anteroom door(s) located between the clean and dirty change rooms not to be opened simultaneously with either the containment laboratory door or the clean change entry door. (Interlock, visual or audible alarms, or protocols are all acceptable means.)
11       Mandatory Anteroom door(s) located between the clean and dirty change rooms not to be opened simultaneously with either the containment laboratory door or the clean change entry door (interlock only).
12     Mandatory Mandatory Interlocked doors, if present, to have manual overrides for emergency exit.
13     Mandatory Mandatory Entry to laboratory zone to be provided with clothing change areas separating personal and laboratory clothing dedicated to that zone (i.e., "clean" change area separated from "dirty" change area).
14     Mandatory Mandatory

Exit from laboratory to be provided with a walk-through shower on the containment barrier (i.e., between “dirty” and "clean" change anterooms).

(CL3 laboratories manipulating organisms, such as HIV, that are not infectious via inhalation, are not required to fulfil this criterion.)
15       Mandatory Entry to laboratory to be provided via anteroom with airtight doors (e.g., inflatable or compression seal); for laboratories using only a Class III BSC biological safety cabinet line, airtight doors are not required.
16       Mandatory Entry to laboratory zone to be provided with a suit change area, a chemical shower on the containment barrier (i.e., between the laboratory and suit change area) and water shower on exit from the zone (i.e., between "dirty" and "clean" change areas); for laboratories using only a Class III biological safety cabinet line, suit change area and chemical shower are not required.
17     Recommended Mandatory Containment laboratories to be located in close proximity to supporting mechanical services to limit the amount of potentially contaminated services.
18     Recommended Mandatory Containment laboratories to be located away from external building envelope walls.
19     Recommended Recommended A laboratory support area to be provided adjacent to the containment facility for all supporting laboratory manipulations.

4.2 Matrix 2
Surface (i.e., floors, walls, ceilings, sealants) Finishes and Casework

Matrix 2 Containment Level Surface (i.e., floors, walls, ceilings, sealants) Finishes and Casework
1 2 3 4
1   Recommended Mandatory Mandatory Doors, frames, casework and bench tops to be nonabsorptive (i.e., the use of organic materials should be avoided).
2   Mandatory Mandatory Mandatory Working surfaces of bench tops to be non-absorptive.
3 Recommended Mandatory Mandatory Mandatory Surfaces to be scratch, stain, moisture, chemical and heat resistant in accordance with laboratory function.
4 Recommended Recommended Mandatory Mandatory Surfaces to provide impact resistance in accordance with laboratory function.
5   Recommended Mandatory Mandatory Surfaces to be continuous and compatible with adjacent and overlapping materials (i.e., to maintain adhesion and a continuous perimeter); wall and floor welded seams are acceptable in level 3 laboratories.
6     Mandatory Mandatory Continuity of seal to be maintained between the floor and wall (a continuous cove floor finish up the wall is recommended).
7     Mandatory Mandatory Interior surfaces to minimize movement of gases and liquid through perimeter membrane.
8 Recommended Mandatory Mandatory Mandatory Interior coatings to be gas and chemical resistant in accordance with laboratory function (e.g., will withstand chemical disinfection, fumigation).
9     Mandatory Mandatory Interior coatings to be cleanable.
10       Mandatory Structural stability to withstand 1.25 times maximum design pressure under supply and exhaust fan failure conditions (i.e., no wall distortion or damage).
11 Recommended Recommended Mandatory Mandatory Bench tops to have no open seams.
12 Recommended Recommended Recommended Recommended Bench tops to contain spills of materials (e.g., with marine edges and drip stops).
13 Recommended Recommended Recommended Mandatory Benches, doors, drawers, door handles, etc. to have rounded rims and corners.
14 Recommended Recommended Recommended Recommended Backsplashes, if installed tight to wall, to be sealed at wall-bench junction.
15 Recommended Recommended Recommended Recommended Reagent shelving to be equipped with lip edges.
16 Recommended Recommended Recommended Recommended Drawers to be equipped with catches, i.e., to prevent the drawer from being pulled out of the cabinet.
17       Recommended Drawers to be of one piece construction.
18 Recommended Recommended Recommended Recommended Cabinet doors not to be self-closing.

4.3 Matrix 3
Heating, Ventilation and Air Conditioning (HVAC)

Matrix 3 Containment Level HVAC
1 2 3 4
1   Recommended Mandatory Mandatory 100% outside air to be supplied.
2     Mandatory Mandatory Directional inward airflow provided such that air will always flow towards areas of higher containment (e.g., ± 25 Pa differential).
3     Mandatory Mandatory Visual pressure differential monitoring devices to be provided at entry to containment laboratory.
4       Mandatory Room pressure differential monitoring lines penetrating the containment barrier to be provided with filters of efficiency equal to that of HEPA filtration.
5     Mandatory Mandatory Alarm (visual or audible) to be provided in the laboratory and outside laboratory area (i.e., to warn others and maintenance personnel) to signal air handling systems failure.
6     Mandatory   Where determined necessary by a local risk assessment, supply air duct to be provided with backdraft protection (i.e., HEPA filter; bubble tight backdraft damper).
7       Mandatory Supply air to be HEPA filtered.
8     Mandatory Mandatory

Supply air system to be independent of other laboratory areas. CL3 supply can be combined with areas of lower containment when provided with backdraft protection (i.e., HEPA filter, bubble tight backdraft damper) downstream from the connection.

(For CL3 laboratories manipulating organisms, such as HIV, that are not infectious via inhalation this criterion is only recommended.)
9     Mandatory Mandatory Supply air system to be interlocked (i.e., fans, dampers, electrical) with exhaust air system, to prevent sustained laboratory positive pressurization.
10     Mandatory Mandatory

Exhaust air to be HEPA filtered.

(CL3 laboratories manipulating organisms, such as HIV, that are not infectious via inhalation are not required to fulfil this criterion.)
11       Mandatory Exhaust air to be passed through two stages of HEPA filtration.
12     Mandatory Mandatory HEPA filters installed into the supply and exhaust system to conform to the requirements of IEST-RP-CC001.3(1).
13       Mandatory Supply HEPA filter housings to be designed to withstand structural change at applied pressure of 2500 Pa [10 in. w.g.].
14     Mandatory   Where HEPA filters are used for backdraft protection in accordance with local risk assessment, supply HEPA filter housings to be designed to withstand structural change at applied pressure of 2500 Pa [10 in. w.g.].
15     Mandatory Mandatory

Exhaust HEPA filter housings to be designed to withstand structural change at applied pressure of 2500 Pa [10 in. w.g.] and to be provided with a method of isolation and decontamination.

(For CL3 laboratories manipulating organisms, such as HIV, that are not infectious via inhalation this criterion is only recommended.)
16     Mandatory Mandatory

Exhaust air system to be independent of other laboratory areas. CL3 exhaust can be combined with areas of lower containment when provided with a HEPA filter upstream from the connection.

(For CL3 laboratories manipulating organisms, such as HIV, that are not infectious via inhalation this criterion is only recommended.)
17     Recommended Mandatory Supply and exhaust systems located outside of containment to be accessible for repairs, maintenance, cleaning and inspection.
18       Mandatory Supply air ductwork that is outside the containment perimeter (e.g., between containment perimeter and HEPA filter or bubble tight backdraft damper) to be sealed airtight in accordance with Sheet Metal and Air Conditioning Contractors National Association (SMACNA) Seal Class A(2).
19     Mandatory   Where backdraft protection is required in accordance with local risk assessment, supply air ductwork that is outside the containment perimeter (e.g., between containment perimeter and HEPA filter or bubble tight backdraft damper) to be sealed airtight in accordance with SMACNA Seal Class A(2).
20     Mandatory Mandatory

Exhaust air ductwork that is outside the containment perimeter (e.g., between containment perimeter and HEPA filter or bubble tight backdraft damper) to be sealed airtight in accordance with SMACNA Seal Class A(2).

(CL3 laboratories manipulating organisms, such as HIV, that are not infectious via inhalation are not required to fulfil this criterion.)
21     Mandatory Mandatory

Airflow control devices and duct sensors to be located downstream of the exhaust HEPA filter and upstream of the supply bubble tight backdraft damper or HEPA filter, or if located upstream, duct penetrations to be sealed in accordance with SMACNA Seal Class A(2).

(CL3 laboratories manipulating organisms, such as HIV, that are not infectious via inhalation are not required to fulfil this criterion.)
22     Mandatory Mandatory

Bubble tight backdraft dampers and HEPA filters to be located in close proximity to the containment perimeter.

(CL3 laboratories manipulating organisms, such as HIV, that are not infectious via inhalation are not required to fulfil this criterion.)

4.4 Matrix 4
Containment Perimeter

Matrix 4 Containment Level Containment Perimeter
1 2 3 4
1 Recommended Mandatory     Autoclave or other acceptable means of waste treatment/disposal to be provided.
2     Mandatory Mandatory

Double-door barrier autoclave with bioseal to be located on containment barrier; body of autoclave to be preferably located outside of containment for ease of maintenance.

(For CL3 laboratories manipulating organisms, such as HIV, that are not infectious via inhalation it is not mandatory that the autoclave be a double-door barrier model.)
3     Mandatory   Barrier autoclave to be equipped with interlocking doors, or visual or audible alarms to prevent both doors from opening at the same time.
4       Mandatory Barrier autoclave to be equipped with interlocking doors, and visual or audible alarms to prevent both doors from opening at the same time.
5     Mandatory Mandatory For materials that cannot be autoclaved (e.g., heat sensitive equipment, samples, film) other proven technologies for waste treatment (e.g., incineration, chemical, or gas) to be provided at containment barrier.
6     Mandatory Mandatory All penetrations to be sealed with nonshrinking sealant at containment barrier.
7     Mandatory Mandatory All conduit and wiring to be sealed with nonshrinking sealant at the containment barrier.
8 Mandatory Mandatory     Windows, if they can be opened, to be protected by fly screens.
9     Mandatory Mandatory Windows positioned on containment barrier to be sealed in place; window glazing material to provide required level of security.
10     Recommended Recommended Observation windows to be installed on containment barrier.

4.5 Matrix 5
Laboratory Services (i.e., water, drains, gas, electricity, and safety equipment)

Matrix 5 Containment Level Laboratory Services (i.e., water, drains, gas, electricity, and safety equipment)
1 2 3 4
1 Mandatory Mandatory     Hooks to be provided for laboratory coats at laboratory exit; street and laboratory clothing areas to be separated.
2 Mandatory Mandatory Mandatory Mandatory Handwashing sinks to be located near the point of exit from the laboratory or in anteroom. Not applicable to CL4 suit laboratories.
3   Recommended Mandatory Mandatory Handwashing sinks to be provided with "hands-free" capability.
4     Mandatory Mandatory BSCs and other primary containment devices to be provided.
5   Recommended     BSCs and other primary containment devices to be provided. Examples for use include procedures with the potential for producing aerosols and those involving high concentrations, large volumes or particular types of agents.
6   Mandatory Mandatory   Emergency eyewash facilities to be provided in accordance with applicable regulations (i.e., ANSI Z358.1-1998(3)).
7   Mandatory     Emergency shower equipment to be provided in accordance with applicable regulations (i.e., ANSI Z358.1-1998(3)).
8     Mandatory   When it is not possible to limit the quantities of hazardous chemicals within the laboratory, emergency shower equipment to be provided in accordance with applicable regulations (i.e., ANSI Z358.1-1998(3)).
9     Mandatory Mandatory Domestic water branch piping serving laboratory area(s) to be provided with backflow prevention, in accordance with CAN/CSA-B64.10-01/B64.10.1-01(4) ,and isolation valve, to be located in close proximity to the containment barrier.
10     Mandatory   Drain lines and associated piping (including autoclave condensate) to be separated from lower containment laboratory areas and to go directly to main building sanitary sewer at point of exit from building (downstream of all other connections).
11       Mandatory Drain lines and associated piping (including autoclave condensate) to be separated from areas of lower containment and to be connected to an effluent sterilization system.
12       Mandatory Drains connected to effluent sterilization to be sloped towards sterilization system to ensure gravity flow; consideration should be given to the installation of valves to isolate sections of piping for in situ decontamination; the effluent sterilization system (e.g., piping, valves, tank) to be heat and chemical resistant consistent with application.
13     Mandatory Mandatory Autoclave condensate drain to have a closed connection. For CL3, open connection is allowable if located within containment barrier.
14     Mandatory Mandatory Drainage traps to be provided to required deep seal depth in consideration of air pressure differentials.
15     Recommended Mandatory Floor drains not to be provided, except when essential (e.g., body shower and animal rooms).
16       Mandatory Plumbing vent lines (including effluent sterilization system) to be provided with filter of efficiency equivalent to that of HEPA and provided with a means of isolation and decontamination.
17     Mandatory  

Plumbing vent lines to be independent of lower containment plumbing vent lines, or combined with lines from lower containment when provided with a filter of efficiency equivalent to that of HEPA upstream from the connection.

(CL3 laboratories manipulating organisms, such as HIV, that are not infectious via inhalation are not required to fulfil this criterion.)
18     Recommended Mandatory Compressed gas cylinder(s) to be located outside the laboratory.
19       Mandatory Laboratory supply gas piping (e.g., carbon dioxide, compressed air) to be provided with backflow prevention.
20     Mandatory Mandatory Portable vacuum pump to be provided in the laboratory. Internal contamination of vacuum pump to be minimized (e.g., HEPA filtration of vacuum line, use of disinfectant traps).
21       Mandatory Compressed breathing air to be provided to positive-pressure personal protective equipment (i.e., for connection to the air hose of suits), equipped with breathing air compressors and back-up cylinders (sufficient for 30 minutes per person); air hose connections to be provided in all areas where suits are worn, including chemical shower and suit change room.
22     Mandatory Mandatory Emergency lighting to be provided.
23     Mandatory Mandatory Life safety systems, lighting, HVAC systems, BSCs, security systems and other essential equipment to be supported with emergency back-up power.
24     Mandatory Mandatory Circuit breakers to be located outside biocontainment area.
25     Recommended Recommended Fluorescent light ballasts and starters to be located outside containment area.
26     Mandatory Mandatory Laboratory to be equipped with a communication system between containment area and outside support area.
27     Mandatory Mandatory System (e.g., fax, computer) to be provided for electronic transfer of information and data from laboratory area to outside laboratory perimeter. (Note: paperwork from the containment laboratory may be removed after appropriate decontamination, i.e., autoclaving, irradiation, microwaving; such practices are generally not recommended for use on a routine basis).
28       Mandatory Work area to be monitored (e.g., closed circuit TV) from outside laboratory perimeter (e.g., security/biosafety office).

References

  1. HEPA and ULPA filters. IEST-RP-CC001.3. Rolling Meadows, IL: The Institute of Environmental Science and Technology, 1993.

  2. HVAC air duct leakage test manual. Chantilly, Virginia: Sheet Metal and Air Conditioning Contractors National Association, Inc., 1985.

  3. American national standard for emergency eyewash and shower equipment. ANSI Z358.1-1998. Arlington, Virginia: American National Standards Institute, Inc., 1998.

  4. Manual for the selection and installation of backflow prevention devices/manual for the maintenance and field testing of backflow prevention devices. CAN/CSA-B64.10-01/B64.10.1-01. Toronto, ON: Canadian Standards Association, 2001.

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Date Modified: 2005-01-05

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