Safety/Risk Groups
Risk Groups
What are Risk Groups and Safety Levels?
Microorganisms are classified into four Risk Groups, and biological laboratories are classified into four corresponding Safety Levels. Risk Group 1 contains non-pathogenic organisms like yeast and E. coli K-12. The majority of iGEM teams use only Risk Group 1 organisms. Some teams use Risk Group 2 organisms. iGEM teams are not permitted to use Risk Group 3 or 4 organisms, or to work in Safety Level 3 or 4 laboratories.
| RG 1 | RG 2 | RG 3 | RG 4 | |
|---|---|---|---|---|
| Examples | E. coli K-12, yeast (S. cerevisiae), Lactobacillus, B. subtilis | Streptococcus, Herpes virus, most mammalian cell lines | Yersinia pestis (black plague), HIV, SARS virus | Ebola virus, Marburg virus, Lassa virus |
| Description | RG 1 organisms do not cause disease in healthy adult humans. | RG 2 organisms can cause disease in humans, but the disease is treatable or preventable. | RG 3 organisms cause serious disease in humans. Treatments and vaccines for these diseases may exist. | RG 4 organisms cause deadly disease in humans, and they can easily travel from one person to another. No treatments or vaccines exist for these diseases. |
| Lab Safety Level | Safety Level 1 | Safety Level 2 | Safety Level 3 | Safety Level 4 |
| Typical Lab Photo |  |
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| Typical Work Area | Open bench | Biosafety cabinet / laminar flow hood | Class 3 biosafety cabinet | Full isolation suits |
How can I find out the Risk Group of my organism?
To determine the Risk Group of an organism, we must consult reliable sources. There is no world-wide standard list of microorganisms and their Risk Groups. Some countries and some cell culture vendors have provided lists, but no single list includes all species, so you might need to check several sources. iGEM recommends three sources for Risk Group information: Canadian PSDS, NIH Guidelines, and DSMZ.
Canadian Pathogen Safety Data Sheets (PSDS)
The Public Health Agency of Canada has published safety data sheets for a wide variety of microorganisms.
To use the PSDS, find your species in the list. Click on the species name to open extensive safety information about that species. Scroll down to "SECTION VII" and look for "RISK GROUP CLASSIFICATION" or "CONTAINMENT REQUIREMENTS".
NIH Guidelines, Appendix B
The NIH Guidelines are a set of rules that govern research on recombinant/synthetic DNA in the United States. Appendix B of the guidelines gives a list of pathogens in Risk Groups 2, 3, and 4. It does not list Risk Group 1 organisms.
Click here to go directly to Appendix B.
DSMZ
"Always check your strain!"
A note about E. coli K-12
Escherichia coli is the most commonly used bacterium in molecular biology. E. coli bacteria naturally live inside the human digestive system. We scientists are used to thinking of E. coli as "safe", but many strains of E. coli cause terrible diseases! Famous strains such as E. coli O157:H7 have caused many deaths. Most disease-causing strains of E. coli are considered Risk Group 2.
E. coli K-12 is a "lab strain". The K-12 strain cannot survive in the human digestive system, and does not produce toxins. K-12 is Risk Group 1, and it may be used in Level 1 laboratories with the most basic safety precautions, such as rubber gloves. Similarly, the strains derived from K-12 (such as DH5alpha, TOP10, etc.) are also Risk Group 1.
Always check your strain!
DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen / German Collection of Microorganisms and Cell Cultures) is a large collection that includes Risk Group information for thousands of different species, strains, and cell lines. It is more difficult to use, but it includes the most species of any source we have found.
- Catalogue Microorganisms -- search bacteria. Enter your genus, species, and/or strain information in the "Search term" box, and click Search. You will receive a long list of results, which may include many different strains and species. Examine the list to find the correct strain, and click on its DSM number to view its catalogue page. The Risk Group is listed in the table.
- Catalogue Human and Animal Cell Lines -- search this catalogue by the same method as for Catalogue Microorganisms.
- Prokaryotic Nomenclature Up to Date is an alphabetical list of all bacterial and archaeal species names with standing in the literature. Find your species in the list, and click on its name to view details. In the table row "Type strain", look for a blue DSM link. If one is present, it will bring you to a DSMZ catalogue page where you can find details about that species. The Risk Group is listed in the table. (If there is no blue DSM link, it means that the DSMZ does not offer that species.) Note: Prokaryotic Nomenclature Up to Date concerns wild organisms, not lab strains. Therefore, it will tell you that E. coli is Risk Group 2. E. coli K-12, or "lab strain" E. coli, is not the same as "wild" E. coli! (For more information, read the orange box on the right.)
Other sources
If you cannot find your species in any of the recommended sources, you must find another reliable source. There may be a paper in the peer-reviewed scientific literature that describes your species. Your country's government might have a list of microorganisms and their Risk Groups.
For help finding a reliable source, you can consult your advisor, or speak to the biosafety authorities of your university. You can also contact <safety AT igem DOT org> for help.
Animals and plants -- what Risk Group are they??
The four Risk Groups only apply to microorganisms (bacteria, fungi, and viruses), with very few exceptions. For animals, plants, and other multicellular organisms, there are different safety guidelines. Your country and your university have their own guidelines governing experiments on animals and plants. You should consult your advisor, your lab manager, or the authorities at your university.
If you are using a part from an animal or plant, you should consider the function of the part, and consider whether it might be dangerous to humans. For example, the gene that codes for Green Fluorescent Protein comes from the jellyfish Aequorea victoria. Therefore, in the strictest interpretation, GFP does not have any associated Risk Group. However, GFP is only a fluorescent protein, absorbing and emitting light, so it is safe for humans and you can use it in a Level 1 laboratory. If you consider a different gene from A. victoria, such as the gene that produces the toxin in its stinging tentacles, you might conclude that you should only use that gene under strict safety conditions!
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