Team:City of London UK/Safety

RIBOTOX

Engineering

Safety

Any piece of work, especially a biological project involving lab work and coming with the future possible implementations in a medical setting, comes with a considerable amount of risk. We have worked diligently to ensure that these risks are reduced considerably, not only for the sake of the iGem regulations and local laws, but also for the sake of ensuring our own safety and aligning with common ethical courtesy. We have applied stringent risk reducing regulations to our laboratory work, hardware and explored the possible safety measures required for our future implementation.

Wet Lab

The live bacteria that we are using are E. Coli JM109 of the K-12 family. They are in risk group 1 and present no risk to humans as they lack certain genes necessary for pathogenic characteristics[1]. The characterisation of our toehold switch constructs took place in a TXTL cell-free kit. This has little risk and so may be safely used outside the lab as a convenient diagnostic tool. Our isothermal amplification also requires no living organisms. We used an RPA kit which is not a biohazard and may be used outside of the lab. However, we kept all of our kit within the safer confines of our laboratory.

Instead of using human plasma samples to test our genetic constructs, we used oligos which we ordered off of IDT. This reduces the need to have unnecessary biohazardous human samples.

As a secondary school team we had further school regulations to follow but we were also given the opportunity to learn safety techniques through the syllabus.

  • Lab access and rules. All labs are locked when out of use and can only be accessed by a responsible teacher with a masterkey. When not in use, our equipment and biological materials are kept in a locked technicians room. We abided by the school laboratory rules and regulations which include a prohibition of eating, drinking, running and throwing. Our microbiology lab and technicians room were clearly signposted so that all persons that were in the vicinity understood the dangers of the area.
  • Responsible individuals. We all understood that any safety queries and issues should be reported to the technician or PI who were always in the lab supervising us.
  • Good microbial technique. We wore microbiology lab-coats at all times as well as wearing gloves, eye protection, trousers and hard, closed footwear. We placed all waste in autoclave bags, with these autoclaved before disposal. All micropipette tips were released directly into virkon which was replaced when the disinfectant was cloudy. Our plates were incubated below human body temperature and were given aerobic conditions.
  • Disinfection and sterilization. We disinfected our work areas with virkon (as opposed to ethanol which is deemed a fire risk) and had the disinfectant close to our work areas. We worked on clean lab mats on top of the lab benches and had a bunsen burner on for sterilization of the environment and equipment.
  • Chemical, electrical and fire safety. We all received training and advice on the interpretation of the warning signs on each chemical and how to use them safely. Protocols and an eye wash kit were in place in case of an emergency. There was a kill switch for all electrical appliances in the room and all members of the team understood the school's fire evacuation procedures.

Hardware

All hardware work was done under teacher supervision. The work that we did also had several safety features, for example, the electromagnets were separated from the solution compartment. On top of that, the contraption comes with a lid. Furthermore, waterproof wiring was used throughout the project. We further mitigated the risk of sparks and fire by working in a laboratory with a fire extinguisher nearby.

Future Implementation

As with any test, there is a chance of a misdiagnosis through false positives or false negatives. This is not a risk unique to our test and so will be mitigated in the common manner of running additional tests.

We will also be taking a blood sample, which although is not uncommon nor dangerous still comes with risks. Some people feel dizzy during and after the test[1].

Refrences

  1. Escherichia coli : mechanisms of virulence. Max Sussman, ed.; Cambridge University Press, 1997. (UCSC Sci Lib Call# QR82.E6 E835 1997)
  2. nhs.uk. 2021. Blood tests. [online] Available at: https: //www.nhs.uk/conditions/blood-tests [Accessed 21 October 2021]