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P2 laboratory ranks third in the safety protection of biological laboratories divided into four safety levels, with the highest being P4, which is a classification of safety levels. Among all types of laboratories now, P2 laboratory is the most widely used biosafety-level laboratory.
1. Determination of main technical indicators
BLS-2 laboratories are mainly used for primary health services, diagnosis, and research. The hazard level of their experimental objects is level II (moderate individual hazard, limited group hazard), which is specifically defined as "pathogens that can cause illness in humans or animals, but generally do not cause serious harm to health workers, groups, livestock or the environment. Laboratory infection does not cause serious disease, has effective ZL and prevention measures, and the risk of transmission is limited." Based on this, the national standard "Technical Specifications for Biosafety Laboratory Construction" stipulates the following technical indicators (static) for BLS-2 laboratories:
1. Cleanliness: no requirements
2. Minimum ventilation times: windows can be opened for ventilation
3. Pressure difference between adjacent rooms connected to the outdoors: no requirements
4. Temperature ℃: 18~27
5. Relative humidity %: 30~70
6. Noise dB (A): ≤60
7. Minimum illumination lx: 300
The above indicators are the minimum requirements for establishing a BLS-2 laboratory. Builders can appropriately increase the corresponding indicators, such as cleanliness, pressure difference, etc., according to actual needs.
2. Building, structure and decoration requirements
Most of the BLS-2 laboratories are now rebuilt in the original buildings. Unlike the newly built BLS-2 laboratories (the national standard requires that the newly built BLS-2 laboratories should be a certain distance away from public places), there is a relative isolation problem between the adjacent rooms. Therefore, when reconstructing the BLS-2 laboratory in the original building, the following issues should be noted:
1. When building a BLS-2 laboratory in a shared building, a door with a lock that can be closed automatically should be set up. If necessary, a buffer area, such as a buffer room, can be set up.
2. If there is no mechanical ventilation system, there should be windows for natural ventilation and insect-proof screens. (Under normal circumstances, there should be a mechanical ventilation system, otherwise, the temperature and humidity indicators are difficult to guarantee).
3. There should be measures to prevent insects, rats and other animals from entering and escaping.
3. Air conditioning, ventilation and purification
In order to meet the temperature and humidity requirements of the BLS-2 laboratory, it is advisable to equip it with a mechanical air conditioning ventilation system. If there are special requirements, additional design requirements can be added, such as adding purification requirements and designing the return (exhaust) air system according to the purification requirements. In this regard, the following issues should be handled well:
1. An air conditioning system with circulating air can be used. If chemical solvents, infectious material operations and animal experiments are involved, a full exhaust system should be used.
2. The biological safety cabinet is connected to the exhaust system.
Level I exhaust ratio, closed connection.
Level II A type exhaust ratio is 30%, which can be discharged to the room or a local exhaust hood can be set up.
Type B1 exhaust ratio is 70%, closed connection.
Type B2 exhaust ratio, closed connection.
3. The location of various equipment in the laboratory should be conducive to the flow of air from the "clean" space to the "contaminated" space, minimizing indoor backflow and leakage (the biological safety cabinet should generally be placed downstream of the indoor airflow, that is, the farthest from the air supply outlet). The exhaust of the biological safety cabinet cannot replace the indoor exhaust.
IV. Drainage
The bacterial sewage should be discharged to the municipal sewage discharge public system after high temperature and high-pressure disinfection or chemical disinfection.
V. Electrical settings
1. A dedicated distribution box should be installed.
2. A leakage detection alarm device should be installed on the power supply.
3. A reliable power connection system should be provided, and its grounding resistance should not be greater than 1Ω.
4. The laboratory entrance should have a text-based light signal display of the laboratory's working status.
5. An emergency luminous evacuation sign should be installed.
VI. Fire protection
The fire resistance level of the building should not be lower than level 2.
VII. Process equipment to be equipped
1. Biosafety cabinet
(1) For operations that produce microbial aerosols or splashes, a Class I biosafety cabinet can be used.
(2) For handling sensitive materials, a Class II biosafety cabinet should be used.
(3) For handling chemical carcinogens, radioactive substances and volatile solvents, a Class II-B biosafety cabinet should be used.
2. High-temperature and high-pressure sterilization and disinfection cabinet.
3. Eyewash station. If necessary, an emergency spray device should be installed.
4. The clean bench does not belong to the biosafety cabinet and cannot be used for biosafety operations.
BLS-2 laboratories are used for preliminary biosafety experiments, and the national standard specifies the corresponding requirements. The establishment of a BLS-2 laboratory should be based on the national standard and determine the construction goals under the premise of ensuring safety. The established laboratory should be a safe and economical laboratory.
VIII. Isolation equipment
1. A biosafety operation device (first or second level) must be set up and regular inspections must be carried out.
2. When using instruments such as grinders, freeze dryers, ultrasonic cell crushers and centrifuges that are prone to generating aerosols to handle recombinants, these instruments must be placed in a safe operation device. However, if the machine already has a device to prevent aerosol leakage, this is not limited.
3. The laboratory must have an autoclave for treating pollutants and sterilizing waste.
IX. Experimental implementation items
1. When conducting experiments, the doors and windows of the laboratory must be closed.
2. After the end of each experiment, the laboratory bench and safety operation device must be sterilized. If contamination occurs during the experiment, it must be sterilized immediately.
3. Waste of biological materials related to the experiment must be sterilized before being discarded. Contaminated instruments must be sterilized by high pressure before cleaning and use or discarding.
4. Do not use your mouth to pipette.
5. Eating, smoking and storing food are prohibited in the laboratory.
6. Wear gloves when operating recombinants to prevent contamination, and wash your hands after the operation and before leaving the laboratory.
7. In all operations, try to avoid the generation of aerosols (for example, when inserting the heated platinum loop and inoculation needle into the culture medium, if a large amount of aerosols are generated, it may cause contamination). Also avoid forcibly ejecting the liquid in the pipette or syringe.
8. When moving contaminated items out of the laboratory, they must be placed in a sturdy and leak-proof container and sealed in the laboratory before they can be transported out.
9. Prevent non-experimental organisms in the laboratory, such as insects and rodents.
10. If other methods are available, avoid using needles.
11. Wear a lab coat in the laboratory and take it off before leaving.
12. People who do not understand the nature of the experiment are prohibited from entering the laboratory.
13. When the experiment is in progress, the entrance of the laboratory should be marked with "P2-level laboratory" and a sign "P2-level experiment in progress" should be hung. The refrigerator and freezer where the recombinant is stored should also be marked in the same way.
14. The laboratory should be cleaned regularly and kept clean. No items unrelated to the experiment should be placed there.
15. Before replacing the HEPA filter in the safety operation device, during regular inspections and when the experimental content is changed, the safety operation device must be sealed and fumigated with 10 grams of formaldehyde per cubic meter for 1 hour to remove contamination.
16. If P1 level experiments are conducted in this level laboratory at the same time, the experimental area must be clearly divided and the operation must be carried out carefully.
17. Other matters specified by the plan host must be followed.
