Difference between revisions of "Team:Duke/Safety"

Our lab, an extension of the Lynch lab, follows the BSL-1 guidelines described as suitable for work involving well-characterized agents that are not known to consistently cause disease in immunocompetent adult humans. The guidelines include washing hands before leaving the laboratory, and careful management of needles and other sharps. Decontamination is strictly followed through autoclaving of solid trash, bleaching of cell waste solutions, and separately disposing of special waste, such as chloramphenicol. In terms of safety equipment, gloves are worn at all times and changed when contaminated. More details on the guidelines can be found at the CDC page and the Duke Safety page.

Safety Training

We received training on general laboratory safety, including personal protective equipment, proper handling of biological and hazardous waste, proper handling of sharps, use of fume hoods, and emergency procedures. We were also trained on general chemical safety, including identification and labeling of hazards, proper storage and use of common chemicals, regulations regarding more hazardous substances, waste disposal procedures, and basic first aid and emergency procedures. In addition, we also learned about fire and life safety, including emergency preparedness and policies.

This training was through various forms of online and in-person training, required by Duke, depending on the safety levels and hazards you are expected to face. For our purposes, we are required to complete online courses for General Laboratory Safety, General Chemical Safety, and Fire/Life Safety. These courses include online modules and a post-course quiz. A description of required training and the training policy is provided at the Duke Safety page.

In addition to these wet lab safety trainings, our members were also certified through the Duke Internal Review Board via the CITI Program to conduct a survey on human subjects. This training included topics such as: .

Local Rules and Regulations

The Occupational and Environmental Safety Office of Duke University covers safety concerns and compliance. We are working directly within the Lynch lab, which are compliant with and in communication with this office, and we have completed the required safety training that they provide. The guidelines are as described in the Duke Biological Safety page.

Organisms and Parts Used

The chassis organisms used for cloning are the E. cloni strain of Escherichia coli by Lucigen, which are in risk group 1 [1]. They were acquired from the Lynch lab.

Project Risks

Duke iGEM's goal is to design a thermostable variant of griffithsin in E. coli for lateral flow assay applications. Our objective is to produce and characterize pieces for implementation into a lateral flow assay.As such, our project mainly deals with modeling, cloning, and heat assays. After our project, the lectin we design would be used in E. coli fermentation. During the duration of our project, and in future experiments related to further characterization of our parts, active HIV/ZIKV virions are NOT necessary. The structure of HIV and ZIKV are well documented. As a result, the specific epitopes on HIV and ZIKV were isolated and used without the risk of infection.

Laboratory Risk Factors and Mitigation

The biggest risk we work with is SYBR Stain, which is a mutagen and irritant, although we use it in small concentrations. Another risk is the antibiotic chloramphenicol, which is strongly anticipated to be a carcinogen by the National Toxicity Program [2]. The E. coli that we use are derived from a K-12 strain and therefore are not particularly dangerous to humans. However, contact with bacteria could still lead to infection or illness, particularly if the bacteria are ingested. In our lab, we wear gloves, sterilize waste, and keep chemicals in cupboards.

Future Risks and Applications

The project is harmless to average real people because the product is a manufactured rapid test, meant to be self contained and non-infectious. The biggest risk of the project would be to healthcare workers administering the rapid tests to HIV infected persons. To mitigate the risk of HIV infection, administrators of the test should wear gloves and properly dispose of biologic materials and tests.

References

1. http://www.lucigen.com/E.-cloni-10G-and-10GF-Chemically-Competent-Cells/#subcat-tabs1
2. http://ntp.niehs.nih.gov/ntp/newhomeroc/roc10/chl_no_appendices_508.pdf