Safety

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During the experiment, we realized that experimental safety is a very important thing. Therefore, we have always followed the following laboratory safety regulations to ensure the safety of each team member and the smooth progress of the experiment.

1. About our project

In our project design, we strictly adhere to all iGEM rules and policies. There are no organisms or activities in this project that exceed the iGEM whitelist or require prior permission from iGEM, and our data collection is unrelated to other humans, with almost no ethical or moral issues.

2. Biosafety rules

According to the infectivity of pathogenic microorganisms and the degree of harm to individuals or groups after infection, pathogenic microorganisms are divided into four categories:

• The first category of pathogenic microorganisms refers to those that can cause very serious diseases in humans or animals, as well as those that have not yet been discovered or have been declared extinct in our country.
• The second category of pathogenic microorganisms refers to those that can cause serious diseases in humans or animals and are relatively easy to spread directly or indirectly between humans, animals and humans, and animals and animals.
• The third category of pathogenic microorganisms refers to those that can cause human or animal diseases, but generally do not pose serious harm to humans, animals or the environment, have limited risk of transmission, rarely cause serious diseases after laboratory infection, and have effective treatment and prevention measures.
• The fourth category of pathogenic microorganisms refers to microorganisms that do not normally cause disease in humans or animals.

The first and second types of pathogenic microorganisms are collectively referred to as highly pathogenic microorganisms.

According to the biosafety protection level of the laboratory for pathogenic microorganisms, and in accordance with the provisions of the national standard for laboratory biosafety, the laboratory is divided into a level 1, a level 2 (which is divided into ordinary type and enhanced type), a level 3, and a level 4.

Experiments on pathogenic microorganisms shall be carried out in laboratories of corresponding grades. The low-grade pathogenic microorganism laboratory shall not engage in the pathogenic microorganism experiment activities that should be carried out in the high-grade pathogenic microorganism laboratory as stipulated in the national pathogenic microorganism catalogue.

The laboratory shall, on the basis of hazard assessment, develop standard operating procedures for specific experimental activities that have a greater impact on safety, including procedures for laboratory biosafety standard operation, testing research and experimental operation, use of relevant instruments and equipment, use of personal protective equipment, laboratory disinfection, disposal of hazardous waste, safe operation of sharp instruments and treatment of experimental emergencies.

The laboratory shall, in accordance with the relevant laws and administrative regulations on environmental protection and the provisions of the relevant departments under The State Council, dispose of waste water, waste gas and other wastes, and formulate corresponding environmental protection measures to prevent environmental pollution. The waste for designing experiments on pathogenic microorganisms must first be subjected to high temperature autoclave treatment. All waste must be sorted and temporarily stored, labeled and not discarded at will.

3. Bsl-2 requirement

• The door of the laboratory should have a window and can be locked, and the door lock and the opening direction of the door should not hinder the escape of indoor personnel.
• A sink should be provided and should be located near the exit of the laboratory.
• A clothing storage or hanging device should be set up at the entrance of the laboratory, and personal clothing and laboratory work clothes can be placed separately.
• The walls, ceilings and floors of the laboratory shall be easy to clean, impermeable to water and resistant to corrosion by chemicals and disinfectants. The ground should be smooth and non-slip, and carpet should not be laid.
• Laboratory cabinets and seats should be stable and their corners should be smooth.
• Laboratory cabinets and their placement should be easy to clean, and the experimental table should be waterproof, corrosion-resistant, heat-resistant and sturdy.
• The laboratory shall have enough space and cabinets for placing laboratory equipment and articles.
• Laboratory equipment, cabinets and articles should be placed reasonably according to the nature and process of the work to avoid mutual interference and cross-contamination, and should not interfere with escape and first aid.
• The laboratory may utilize natural ventilation. If mechanical ventilation is used, cross contamination should be avoided.
• If there are Windows that can be opened, install mosquito repellent screens.
• Unnecessary reflection and strong light should be avoided in the laboratory.
• If handling irritating or corrosive substances, eye washing facilities should be set up within 30 m, and emergency spray equipment should be set up if necessary.
• If handling toxic, irritating and radioactive volatile substances, appropriate negative pressure exhaust cabinets should be equipped on the basis of risk assessment.
• If highly toxic and radioactive substances are used, corresponding safety facilities, equipment and personal protective equipment shall be equipped, which shall comply with relevant national and local regulations and requirements.
• The door of the main entrance of the laboratory and the door of the laboratory where the biosafety cabinet is placed shall be automatically closed; The door at the main entrance of the laboratory shall have access control measures.
• There shall be conditions for the storage of spare materials outside the laboratory work area.
• Eye washing devices shall be provided in the laboratory work area.
• The laboratory or the building in which it is located shall be equipped with autoclaves or other appropriate disinfection and sterilization equipment based on a risk assessment.The laboratory or the building in which it is located shall be equipped with autoclaves or other appropriate disinfection and sterilization equipment based on a risk assessment.
• Biosafety cabinets shall be provided in laboratories where samples of pathogenic microorganisms are handled. The biosafety cabinet shall be installed and used according to the design requirements of the product. If the exhaust air of the biosafety cabinet is circulated indoors, the room should have ventilation conditions; If a biosafety cabinet is used that requires duct exhaust, it should be discharged through a duct independent of the other public ventilation systems in the building.
• There shall be a reliable supply of electricity. When necessary, important equipment (such as incubators, biosafety cabinets, refrigerators, etc.) should be equipped with backup power.

4. For bsl-2 lab, the configuration and operation of laboratory facilities and equipment shall follow the following guidelines:

(1) Use at all times a properly maintained biosafety cabinet, preferably a secondary biosafety cabinet, or other suitable personnel protection facilities or physical containment devices.

Experimental procedures to determine the possible formation of infectious aerosols or spillages include centrifuging, grinding, homogenizing, violent shaking or mixing, ultrasonic cracking, opening the container containing the infectious agent (the pressure inside the container may not be consistent with atmospheric pressure), animal nostril inoculation, and collection of infected tissue from animals or embryonic eggs.

When a high concentration or large volume source of infection is involved, if a centrifuge with a sealed rotor or a safety cover is used, if the rotor or safety cover is only opened in the biosafety cabinet, the centrifuge can be centrifuged in an open laboratory.

(2) When microorganisms must be handled outside the biosafety cabinet, facial protection measures (mirror, mask, mask, or other anti-splash device) should be taken to prevent infectious sources or other harmful substances from splashing or spilling on the surface.

(3) In the laboratory, special protective clothing, gown, smock or uniform must be used. When personnel go to non-laboratory areas, protective clothing must be left in the laboratory. Protective clothing can be handled in the laboratory or washed in the laundry room, but not taken home.

(4) Wear gloves when possible contact with potential sources of infection, contaminated surfaces or equipment. Two pairs of gloves are more appropriate. Gloves that are clearly contaminated should be disposed of and removed when the work on the source of infection is completed or the gloves are damaged. Disposable gloves do not need to be cleaned, cannot be reused, cannot be used to contact "clean" surfaces (keyboards, telephones, etc.), and should not be worn outside the laboratory. Have latex gloves with talcum powder. Wash your hands after removing gloves.

(5) The surface of the test bench should be waterproof, heat resistant, organic solvent resistant, acid and alkali resistant and durable in the workbench and other chemical substances for disinfection of facilities; There should be an eye irrigation device; If the laboratory has Windows facing the outside, window screens should be installed to prevent flies.

(6) When installing the biosafety cabinet, it is necessary to take into account the ventilation and exhaust of the room, which will not cause the biosafety cabinet to operate beyond normal parameters. The biosafety cabinet should be far away from the door, away from the window that can be opened, away from the walking area, and away from other equipment that may cause wind pressure confusion, to ensure that the airflow parameters of the biosafety cabinet are within the effective range.

5. Personal protective measures

(1) The laboratory director shall prohibit or restrict access to the laboratory while working on the source of infectious disease. In general, people who are susceptible or who will have serious consequences if infected are not allowed to enter the laboratory, for example, people with immune deficiencies or immunosuppression, who are at increased risk of infection. The laboratory director has the ultimate responsibility for assessing each situation and deciding who can work in the laboratory.

(2) Laboratory personnel receive appropriate immunizations or tests related to the source of the disease being treated or to be treated in the laboratory (e.g., hepatitis B immunization or TB skin tests); Where appropriate, basic serum samples appropriate for use by laboratory personnel and persons at risk are collected and stored, depending on the source of the disease being treated. Other serum samples should be collected regularly depending on the function of the disease source or facility being treated.

(3) Biosafety procedures shall be included in the standard operating procedures or biosafety manual specially developed by the laboratory Director for laboratory personnel. For persons with special risks, it is required to read and follow the work and procedures.

(4) The laboratory Director ensures that the laboratory and its supporting staff receive appropriate training, including possible risks associated with the work, necessary measures to prevent exposure, and procedures for exposure assessment. When changes in procedures are necessary, the personnel concerned must update their knowledge annually and receive additional training.

(5) In the presence of an exogenous source of disease, a biohazard sign shall be posted at the entrance to the laboratory, and the following information shall be displayed: the source of the disease, the biosafety level, the immunization requirements, the name of the researcher, the telephone number, the personal protective equipment that must be worn in the laboratory, and the procedures required for leaving the laboratory.

(6) Do not eat, smoke, wash glasses and make up in the work area. Food and daily necessities are not allowed in the work area. In the laboratory, people who wear hidden glasses also need to wear masks or face masks. Food should be stored in a dedicated cabinet or refrigerator outside the work area.

(7) Can not use mouth pipetting, only mechanical device pipetting; Develop safe use of sharp instruments. For contaminated sharp instruments, must always maintain a high degree of vigilance, including needles, syringes, slides, sampler, capillaries, scalpels; If you can use other tools, do not use sharp tools. When possible, use plastic instead of glass. Broken glassware can not be handled directly by hand, must be handled with other tools, such as brushes and dustpans, clips or tweezers. Containers containing contaminated needles, sharps and broken glass should be disinfected in accordance with regulations before they are dropped.

(8) All operating processes should be as careful as possible to avoid sputtering and aerosols; Disinfection should be carried out at least once a day, and tabletop disinfection should also be carried out in time when the living body spills. Spillage or accidental exposure to an apparent source of infection should be reported immediately to the laboratory director. Conduct appropriate medical evaluation, observation, treatment, and keep written records.

(9) All cultures, stores and other specified wastes shall be disinfected by feasible disinfection methods, such as autoclaving, prior to release. Materials transferred to the nearest laboratory for disinfection should be placed in durable, leak-proof containers, sealed and shipped out of the laboratory, and their packaging should comply with the relevant national regulations.