New Coronavirus Laboratory Biosafety Guidelines Second Edition

Source: Institute of Viral Diseases Release time: 2020-01-24
According to the current information on the biological characteristics, epidemiological characteristics, pathogenicity, and clinical manifestations of the new coronavirus, the pathogen is temporarily managed according to the second category of pathogenic microorganisms in the classification of pathogenic microorganisms.

  1. Biosafety requirements for experimental activities

(1) Virus culture: refers to operations such as virus isolation, culture, titration, neutralization test, live virus and its protein purification, virus freeze-drying, and recombination experiment for producing live virus. The above operations should be carried out in a third-level biosafety laboratory. Use virus culture to extract nucleic acid. The addition of lysing agent or inactivating agent must be carried out under the same level of laboratory and protective conditions as virus culture. After adding lysing agent or inactivating agent, it can be compared with the protection of uncultured infectious materials. Level to operate. Before carrying out relevant activities, the laboratory shall report to the National Health Commission for approval and obtain the qualifications for carrying out the corresponding activities.

(2) Animal infection experiments: refer to experimental operations such as infecting animals with live viruses, sampling infected animals, handling and testing infectious samples, special inspections of infected animals, and disposal of infected animal excrement, which should be performed in a third-level biosafety laboratory. Before carrying out relevant activities, laboratories shall report to the National Health Commission for approval and obtain the qualifications for carrying out corresponding activities.

(3) Operation of uncultured infectious materials: Refers to the virus antigen detection, serological detection, nucleic acid detection, biochemical analysis, and the inactivation of clinical samples before inactivation of uncultured infectious materials by reliable methods. Work and other operations shall be carried out in the biosafety level two laboratory, and the personal protection of the biosafety level three laboratory shall be adopted.

(4) Operations of inactivated materials: nucleic acid testing, antigen testing, serological testing, biochemical analysis and other operations that are carried out after inactivating infectious materials or live viruses by reliable methods shall be carried out in the secondary biosafety laboratory. Other operations that do not contain pathogenic live viruses, such as molecular cloning, can be performed in a biosafety first-level laboratory.

  1. Transport and management of pathogens and specimens

(A) Domestic transport: novel coronavirus strains or transport packaging other potentially infectious biological material classification is A class, UN number corresponding to UN 2814 , packaging compliance with ICAO document Doc9284 “Safe Transport of Dangerous Goods by Air Technical Instructions,” the PI602 classification packaging requirements; environmental samples belong to B class, UN number corresponding to UN 3373 , packaging compliance with ICAO document Doc9284 “safe transport of dangerous goods by Air technical Instructions,” the PI650 classification packaging requirements; through other transportation service may refer to the above criteria package.

The transportation of new coronavirus strains or other potentially infectious materials shall be carried out in accordance with the “Administrative Regulations on the Transportation of Highly Pathogenic Microorganisms (Virus) Species or Samples that Can Infect Humans” (formerly Ministry of Health Order No. 45 ). “.

(2) International transportation: If the new coronavirus strain or sample is transported internationally, the packaging should be standardized, and the relevant procedures should be completed in accordance with the “Regulations on the Administration of Sanitary and Quarantine of Entry and Exit Special Articles”, and meet relevant national and international requirements.

(3) Management of strains and samples: The new coronavirus strains and related samples should be managed by dedicated personnel, accurately record the source, type, quantity, and number registration of the strains and samples, take effective measures to ensure the safety of the strains and samples, and take strict precautions Misuse, malicious use, theft, robbery, loss, leakage and other incidents occurred.

  1. Waste management

(1) Laboratories that carry out experimental activities related to the new coronavirus should formulate waste disposal procedure documents and sewage and sewage treatment operating procedures.

(2) All hazardous wastes must be completely and compliantly labeled in accordance with standardized containers and labeling methods.

(3) Properly trained personnel should use appropriate personal protective equipment and equipment to handle hazardous waste.

(4) Waste treatment measures: Waste treatment is a key link in the control of laboratory biosafety. To effectively and safely handle infectious waste, it is necessary to fully grasp the classification of biosafety waste and strictly implement the corresponding treatment procedures.

1 . Treatment of waste liquid: The waste liquid produced in the laboratory can be divided into ordinary sewage and infectious waste liquid.

( 1 ) Ordinary sewage is generated in hand-washing sinks and other equipment. Such sewage should be collected separately, discharged into the laboratory water treatment system, and discharged after treatment reaches the standard.

( 2 ) Infectious waste liquid is the waste water generated during the experimental operation. It is treated by chemical disinfection or physical disinfection, and the disinfection effect is verified to ensure complete inactivation.

( 3 ) The staff shall dispose of the waste in time and shall not take the waste out of the experimental area.

2 . Solid waste treatment

( 1 ) Solid waste is collected separately. The collection container of solid waste should have the characteristics of not easy to break, leakproof, moisture-resistant, heat-resistant, and sealable. Infectious garbage in the laboratory is not allowed to be accumulated and stored, and should be sterilized by pressure steam in time. Before disposal, waste should be stored in a designated safe place in the laboratory.

( 2 ) Small solid wastes such as tissue samples, consumables, personal protective equipment, etc. need to be sterilized by pressure steam, and then removed from the laboratory along the waste channel.

( 3 ) Large solid wastes such as HEPA filters should be disinfected in situ by professionals and then put into safe containers for disinfection and sterilization. Items that cannot be steam sterilized, such as electronic equipment, can be disinfected with ethylene oxide fumigation.

( 4 ) The solid wastes removed from the laboratory after disinfection and sterilization are collectively handed over to the solid waste treatment unit for disposal.

( 5 ) During the experiment, if sharps (including needles, knives, metal and glass, etc.) are used, they should be directly discarded in the sharps box, autoclaved, and then treated uniformly.

(5) Establish waste treatment records: regularly leak detection and replacement of the laboratory exhaust HEPA filter, regularly monitor the treated sewage, and use biological indicators to monitor the effect of pressure steam sterilization.

  1. Mistakes or accidental handling of laboratory biosafety operations

(1) The new coronavirus strains or other potentially infectious materials contaminate the operating platform of the biological safety cabinet and cause limited pollution: use a disinfectant with an effective chlorine content of 0.55% . The disinfectant needs to be used and used within 24 hours. The content of available chlorine in the following content refers to this concentration.

(2) Fragmentation or overturning of virus-containing culture vessels causes laboratory pollution: keep the laboratory space closed to avoid the spread of pollutants, and use 0.55% effective chlorine disinfectant towels to cover the contaminated area. When necessary (when a large amount of spilling), peracetic acid can be used to heat the fumigation laboratory, the dose is 2g/m 3 , fumigation overnight; or 20g/L peracetic acid disinfectant is sprayed with an aerosol sprayer, the dosage is 8mL/m 3 , the effect is 1 ~ 2 hours; if necessary, or fumigate with potassium permanganate – formaldehyde: potassium permanganate 8g/m 3 , put it in a heat-resistant and corrosion-resistant container (terracotta pot or glass container), and then add formaldehyde ( 40% ) 10mL/m 3. Fumigation for more than 4 hours. The indoor humidity is 60-80% during fumigation .

(3) To clean up pollutants, strictly follow the requirements for safe operation of live viruses, use pressure steam sterilization, and conduct laboratory ventilation to prevent secondary hazards.

https://pubs.acs.org/doi/full/10.1021/jacs.0c04253?ref=vi-chemistry_coronavirus_research