1. Lyme Disease: Testing for Borrelia burgdorferi By Maddie Tango

2. Purpose Lyme disease: Caused by bacteria Borrelia burgdorferi Spread to humans by deer ticks (aka black-legged ticks) Originates in rodents Tick bites infected rodent, then bites human  human contracts Lyme Adult Female Deer Tick Source: https://identify.us.com/idmybug/ticks/ti ck-images/deer-tick-adult-female.html

3. Purpose Symptoms Flu-like (fever, chills, body aches, headache) Joint pain Inflammation of membranes surrounding brain Temporary paralysis on one side of face Numbness Weakness Swollen glands Arthritis 50% of patients: classic “bull’s-eye rash” at site of bite Bull’s-Eye Rash Source: http://www.cdc.gov/lyme/signs_sympt oms/index.html

4. Purpose Problem How does a person know if a tick has Lyme disease? Current solution Send tick in to lab for testing (expensive, takes time) Precautionary antibiotics (bad if not needed; damages bacterial flora in gut) My solution Genetically modify bacteria to glow in the presence of Borrelia burgdoferi Find a tick? Drop it in a solution with this GM bacteria to see if it carries Lyme

5. Competing Technologies Send into lab (expensive, takes time) PCR (not always accurate; easy to contaminate) Grow the bacteria in the tick’s gut in order to better detect Borrelia burgdorferi Putting antibodies in tick’s gut  attaches to Borrelia if present  glows (this is not specific to Borrelia burgdorferi, though) Tests on human blood Test for certain antibodies that would be present if Borrelia burgdorferi is also present

6. The Design Bacteria would be genetically modified to glow in the presence of Borrelia burgdorferi and put in a liquid solution Quorum sensing: all bacteria produce unique chemical to let other bacteria know their presence Take receptor gene to that chemical from Borrelia burgdorferi and insert it into the new bacteria  sense presence of Borrelia burgdorferi Connect receptor to GFP gene  glow in presence of chemical released by Borrelia burgdorferi

7. The Design

8. Expected Results Borrelia burgdorferi PresentGlow Green 11 00 If the chemical produced by Borrelia burgdorferi is present (and therefore Borrelia burgdorferi is present), the bacteria will glow green. If it is not present, it will not glow green.

9. Advantages Quick test  Sooner diagnosis Easy Eliminates use of unneeded antibiotics Potential Problems Mutations Working with Borrelia burgdorferi during testing  could contract disease Borrelia burgdorferi Source: http://mph720davis.blogspot.com/2010_07_01_archive.html

10. Testing Put Borrelia burgdorferi in presence of bacterial solution and see if it glows Glowing Bacteria Source: https://andydvorak.net/articles/photoshop/glowing-bacteria

11. Sources (excluding photos) http://labtestsonline.org/understanding/analytes/lyme/tab/sample/ http://lymedisease.org/news/lyme_disease_views/culture-test.html http://www.cdph.ca.gov/HealthInfo/discond/Documents/TickTestQandA.pdf http://www.lymediseaseassociation.org/index.php/about-lyme/medical/symptom-list http://www.lymediseaseassociation.org/index.php/about-lyme/tick-vectors/photos http://www.maine.gov/dhhs/mecdc/infectious-disease/epi/vector-borne/lyme/lyme- faq.shtml http://www.maine.gov/dhhs/mecdc/infectious-disease/epi/vector-borne/lyme/lyme- faq.shtml http://www.mayoclinic.org/diseases-conditions/lyme-disease/basics/symptoms/con- 20019701 http://www.mayoclinic.org/diseases-conditions/lyme-disease/basics/symptoms/con- 20019701 http://www.tickencounter.org/tick_testing http://www.wwhd.org/TLD_CD/ticks.htm