BSL-2 laboratory management system of carrier Engineering Center

This management system specifies the purpose, goal and system of safety management of BSL-2 Laboratory of carrier Engineering Center of Beijing brain science and brain like Research Center (CIBR). This system is also applicable to BSL-1 laboratory.
The standard operating procedures specified in this system are applicable to all BSL-1, BSL-2 laboratories and relevant laboratory staff in the center, including but not limited to CIBR staff, students, visiting scholars, further education and internships, temporary visitors and non CIBR logistics support personnel.
At present, the main pathogens allowed to operate in the virus vector center include recombinant adeno associated virus (AAV), recombinant lentivirus (LV), rabies virus (RV), vesicular stomatitis virus (VSV), pseudorabies virus (PRV), herpes simplex virus (HSV), cytomegalovirus (CMV). If it is necessary to operate other viruses, it shall be reported to the person in charge of the laboratory and the relevant superior competent department for filing before approval.
Standard operating procedures
1. Access control
Only trained and authorized personnel can enter BSL-2 laboratory.
The laboratory door shall be pasted with the international general biological hazard warning signs, indicating the biosafety level, laboratory principal and emergency contact number.
The laboratory door should remain closed.
Items not related to the laboratory shall not be brought into the laboratory.
2. Personal protection
When working in the laboratory, appropriate personal protective equipment (such as protective clothing, gloves, masks, hats, etc.) must be used. No one is allowed to enter the working area of BSL-2 laboratory directly wearing personal clothes or ordinary laboratory clothes. After changing the buffer parts, he must enter the laboratory with the laboratory clothes of BSL-2 laboratory.
In order to prevent eyes or face from being hurt by splashes, bumps or ultraviolet rays, goggles, masks or other protective equipment shall be worn when necessary.
Do not wear toes exposed shoes in the laboratory.
It is strictly prohibited to leave BSL-2 laboratory wearing special protective clothing, gloves and other protective equipment.
Never touch the door handle, light switch, telephone, computer and other public facilities with hands wearing gloves that may have been contaminated.
Gloves shall be replaced immediately in case of damage.
In case of spilling or overflowing of pathogenic microorganism in BSL-2 laboratory, the operator must take emergency measures immediately and report to the safety administrator as soon as possible.
It is forbidden to eat, drink, smoke, make-up and handle contact lenses in BSL-2 laboratory.
It is forbidden to store food, beverage and other non experimental items in BSL-2 laboratory.
3. Operation specification
The laboratory is divided into polluted area, semi polluted area and clean area, which are distinguished by color identification. The above approved class III pathogens can only be operated in the contaminated area, and only the contaminated materials sealed in the sterilized containers can be obtained in the semi polluted area. The cleaning area shall not operate the reproducible and active pathogens.
The operation of any active third class pathogens should be carried out in the class II biosafety cabinet. Open test bed is not allowed to operate such pathogens.
All BSL-2 laboratory guidelines must be followed when operating active class III pathogens in a BSL-2 laboratory.
When operating the third type of active pathogens outside the level II biosafety cabinet (for example, in the process of putting the cell culture bottle inoculated with virus into the incubator), the principle of double-layer protection shall be observed.
The laboratory should be kept clean and tidy. Only the instruments and articles currently in use can be placed on the test bench. Consumables shall be stored in designated cabinets or shelves and taken as needed.
It is forbidden to use mouth to suck liquid; auxiliary liquid transfer device must be used.
Syringes and needles can only be used when they are absolutely needed. The used needles must not be capped or bent again. They must be placed in a special sharp tool box.
Try to use plastic or experimental equipment that is not easy to be damaged. Glass products can only be used when necessary in the experimental steps (such as using micro slides, etc.).
The instruments or equipment of BSL-2 laboratory can only be taken out of the laboratory with the approval of the laboratory administrator and the person in charge. If the instrument cannot be thoroughly sterilized, it is not recommended to take it out of the BSL-2 laboratory. The scrapped instruments can only be disposed after being disinfected by appropriate means.
Aerosol should be avoided during operation. Centrifugation should be carried out in the turning head of anti aerosol. Turn around is only allowed to open, load and unload in the biosafety cabinet. Suction head and pipette with filter membrane must be used.
All test tubes, centrifuges, reagent bottles, culture bottles, Petri dishes, etc. must be clearly marked and can only be placed in the incubator, refrigerator or freezer in the BSL-2 laboratory.
The third kind of active pathogens shall not be taken out of the laboratory unless approved by the person in charge of the laboratory.
The center's biosafety Executive Committee reviews the biosafety registration at least once a year.
It is not recommended to bring books or experimental records into BSL-2 laboratory. It is suggested that the experimental records made in BSL-2 laboratory should be sent out to the laboratory in the form of scanning and photographing. It is forbidden to take the paper directly out of the polluted area. If it must be taken out, it is recommended to put it into the plastic sheet and disinfect the surface.

4. Use biosafety cabinet
Based on the principle of aerodynamics, the class II biosafety cabinet can not only provide protection for operators, but also provide protection for operating samples. When using the biosafety cabinet, the following regulations must be observed:
The biosafety cabinet shall be operated for at least 15 minutes before operation.
Clean and disinfect the biosafety cabinet first, and then put the experimental supplies in the cabinet.
Only the necessary materials and supplies for this experiment are placed in the biosafety cabinet. In order to avoid cross contamination, the laboratory supplies should be disinfected with 70% alcohol before being taken into the biosafety cabinet.
When operating the third type of active pathogens in the biosafety cabinet, all articles, equipment and instruments in the cabinet shall be treated as pollutants.
The biosafety cabinet shall be cleaned and disinfected after the completion of the work. It shall be disinfected by ultraviolet radiation for 1 hour every day (automatically cut off after 1 hour) to strengthen the disinfection effect.
5. Good housekeeping
In order to minimize the potential risks caused by unclean working environment, microorganism and improper operation of laboratory protective clothing, the hygiene plan should be strictly followed. The hygiene plan includes the principles of cleaning and disinfection of hands, biosafety cabinets, laboratory equipment, floors, rooms and protective clothing.
The staff of BSL-2 laboratory shall be responsible for cleaning the used worktops and materials after work. The students on duty shall clean the ground in the working area every day, once a week, and once a month.
6. Waste disposal
All the experiments related to the above viruses will be carried out in BSL-2 laboratory. The liquid pollution waste generated shall be collected in a special covered closed waste liquid tank. When the volume reaches 3 / 4, add an appropriate amount of chlorine containing disinfectant (such as calcium hypochlorite solution, etc., and the effective chlorine needs to be greater than 2000mg / L) for 2 hours, and then use 75% alcohol to treat the surface of the waste liquid tank, and then remove it from BSL-2 Laboratory for dumping In the laboratory waste water tank.
Solid pollution wastes (such as pipette, culture dish, culture bottle, etc.) shall be collected in the garbage bag that can be sterilized by high pressure. The damaged wastes (pipette gun head, sharp tool) shall be collected in the Yellow sharp tool box with a cover and puncture resistance. After sealing and using 75% alcohol to treat the surface, they shall be removed from the BSL-2 laboratory and immediately placed in the sterilization pot for high pressure sterilization. After that, they can be treated as ordinary Laboratory waste disposal.
Other non disposable materials involving virus operation (such as ultracentrifuge sleeve, centrifuge tube, etc.) shall be soaked with 75% alcohol; surfaces that may involve overflow or splashing of virus liquid shall be wiped with 75% alcohol.
7. Exit BSL-2 Lab
Before leaving BSL-2 laboratory, the laboratory personnel shall confirm that the experimental area and articles have been cleaned and the wastes generated by their own experiments have been properly disposed; the last laboratory personnel leaving the laboratory at the end of work shall be responsible for: - verify that all lights and doors have been closed and all equipment not set to continue to operate have been closed. In case of any abnormal conditions and events, the laboratory personnel shall report to the emergency response team on duty in time.
8. overtime work
The normal working hours of CIBR are 9:00-17:00 from Monday to Friday. If not necessary, please try to avoid working in BSL-2 laboratory outside normal working hours, including weekends and holidays. If it is necessary to work overtime due to work, the laboratory administrator or the person in charge of safety shall be informed in advance, and the access record shall be truthfully registered.

9. Service and maintenance
Before any maintenance work, the laboratory safety director must disinfect the corresponding area / equipment. The maintenance and repair personnel shall be informed of the potential biological hazards in the work area. The staff shall wear appropriate protective equipment (disposable protective clothing, etc.) and carry out the work under the supervision of qualified staff of BSL-2 laboratory.
10. Fire, earthquake and other emergencies
The experimental operation must comply with the fire regulations. Fire extinguishers (CO2 extinguishers) in the BSL-2 laboratory may be used when handling small scale fires. The laboratory is equipped with a heat and smoke detection system to detect undetected fires or fires occurring outside the working day.
If the alarm in the building sounds or the red strobe lights up, the staff in BSL-2 laboratory must evacuate immediately (after closing the window of the biosafety cabinet). The safety of personnel is more important than any experiment! Personnel can evacuate through buffer room, air lock room or emergency exit. If the door cannot be opened through the magnetic lock, the breakable glass plate shall be used for evacuation.
11. Contingency plans for accidents, splashes, spills, etc
In case of accidents such as personal injury, other personnel must report to the emergency response team, the biosafety supervisor and the laboratory on duty immediately. The emergency team shall supervise the cleaning process of spills and decide whether to take further safety measures and whether to take medical measures.
In BSL-2 laboratory, a large amount of liquid containing virus overflows or splashes. In this case, the spilled liquid or splashed surface shall be quickly wiped with absorbent paper, and then the spilled or splashed area shall be wiped with 75% alcohol or chlorine containing disinfectant, and the wipes of absorbent paper shall be collected in the red high-pressure sterilization bag and sterilized. If the pollution is serious, it is necessary to fumigate the whole BSL-2 laboratory with formaldehyde or hydrogen peroxide.
12. training
All the experimenters must be trained and qualified before they can work in BSL-2 laboratory. Working experience in other BSL-2 and above protection level does not mean that access right has been obtained, but training is still required before entering the BSL-2 Laboratory of CIBR. The training contents mainly include necessary safety operation skills and how to deal with emergencies. The training methods include BSL-2 laboratory operation, biosafety education lectures and hands-on training. The training records of the experimenter and the approval documents for the use of the laboratory shall be kept on file.
Every year, the center needs to hold a biosafety lecture applicable to the central laboratory. The authorized experimenter must attend the lecture before the authorization. The authorized person also needs to review the relevant knowledge awareness authorization to continue to take effect. All qualified staff and trainees of BSL-2 laboratory must attend and implement the sign in system, and the sign in record shall be kept by the person in charge of BSL-2 laboratory.
Mutual trust, sincerity and respect should be the core values of BSL-2 laboratory staff. Therefore, it is necessary and obligatory for each experimenter to comply with the laboratory regulations, so that they can trust and be trusted by their colleagues in daily work and emergencies.
13. Occupational health assessment
Occupational health assessment shall be carried out before all training work and regular training after the training is qualified. Please refer to the guidelines for occupational health assessment for details.
It is not recommended to work in BSL-2 Laboratory for people with immunosuppression problems, respiratory diseases, and allergies to latex, animals or disinfectants. Pregnant female workers working in BSL-2 laboratory need special attention. It is not recommended to operate toxic or infectious pathogens and reagents that may affect the fetus.

14. Disciplinary action
Violation of biosafety guidelines may not only harm the individual health of the violator, but also endanger colleagues and the environment. For serious and repeated violations of biosafety guidelines, BSL-2 laboratory safety responsible person shall report to the center and take punishment.
Serious violations of this biosafety guideline include, but are not limited to, the following:
-Ignore laboratory and personal hygiene regulations. If you eat or drink in the laboratory, do not disinfect every time you use the biosafety cabinet. Do not wash hands or disinfect before leaving the laboratory.
-Do not comply with public obligations, such as no high-pressure waste, no add consumables and do not participate in the routine cleaning work of the laboratory.
-Do not comply with the regulations of the visitors, such as allowing friends, family or service maintenance personnel to enter the laboratory without the permission of the safety responsible person of BSL-2 laboratory, or leaving the visitors away from the supervision in the laboratory.
-Do not handle in time and report accidents or spills of infectious materials to the emergency response team.
-Enter the laboratory during shutdown for safety reasons.