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MRSA DiagnosticsKaitlyn Tingley & Amanda BukhtiaLawrence Technological University
Advisors: Dr. Li, Dr. Nasir, Dr. Bou-Akl and Dr. Auner
What is MRSA?● Methicillin-resistant
Staphylococcus aureus(MRSA)
● An antibiotic resistant staph infection
● Enter the bloodstream through open wounds leading to death
http://www.cdc.gov/mrsa/tracking/index.html
1
Need Statement
“Develop a quick and cost effective method to detect if a patient has
(MRSA)”
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Market Value
Stakeholders•Hospitals•Nursing Homes•Schools•Patients
Market Value•In VA hospitals alone the market value is 30.94 million
http://www.mayoclinic.org/diseases-conditions/mrsa/basics/definition/con-20024479http://www.aaos.org/news/aaosnow/mar09/clinical13.asp
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Current Diagnostic Methods4
http://laboratory-manager.advanceweb.com/Archives/Article-Archives/MRSA-Screening-Culture-or-Molecular.aspxhttp://ac.els-cdn.com/S0956566311005197/1-s2.0-S0956566311005197-main.pdf?_tid=55edafa6-5648-11e4-beaa-00000aacb35f&acdnat=1413583120_a5737397e845f21a215313800649bf76http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1287783/ http://www.rapidmicrobiology.com/test-method/methicillin-resistant-staphylococcus-aureus-mrsa-detection-and-identification-methods/http://www.biomedcentral.com/1471-2180/6/83https://workspace.imperial.ac.uk/opticalandsemidev/public/Publications/LAWES.pdfhttp://www.aist.go.jp/aist_e/latest_research/2012/20121121/20121121.html
Diagnostic Steps
1. Separation2. Concentration3. Detection
Lowy, Franklin. "STAPHYLOCOCCUS AUREUS INFECTIONS." New England Journal of Medicine (1998): 520-32. Print. http://www.medicinenet.com/staph_infection/page5.htm#what_is_antibiotic-resistant_staph_aureus
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Biological InteractionsPenicillin-Binding Protein 2a
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http://jb.asm.org/content/187/5/1815.fullhttp://www.britannica.com/EBchecked/topic/27783/antibody
MRSA
AntiPBP2a
MRSANonMRSA
Anti PBP1AntiPBP2a
Design Concept
.
71
Slide 8
1 Finish NotesKaitlyn Tingley,
Project Tasks
1. Device Fabrication 2. Antibody Immobilization3. Simulation to determine flow parameters 4. Device Testing
82
Slide 9
2 Finish NotesKaitlyn Tingley,
Electrode Fabrication
• Substrate Material: Glass (slide)
93
Slide 10
3 Finish NotesKaitlyn Tingley,
Flow Channel Fabrication
● Soft Lithography
104
Slide 11
4 Finish NotesKaitlyn Tingley,
Antibody Immobilization and Device Bonding11
1. Antibody Immobili-zation 2. Device Bonding
Device Testing12
Adapted from Dr. Nasir
Inlet Outlet
PDMS
Glass
Au Electrodes Non-MRSAMRSA
5
Slide 13
5 Finish NotesKaitlyn Tingley,
Testing Parameters
• Staph SolutionMSSA (Control)MRSA(Variable)
• Input: Staph Solution
• Output: Impedance
1367
Slide 14
6 Finish NotesKaitlyn Tingley,
7 Double check last sentenceKaitlyn Tingley,
Validation Testing
http://www.inphotonics.com/raman.htmHarper, Chem Commun,2012, 48(75):9412-4
Raman Spectroscopy
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Blue is MRSARed is Non-MRSA
Expected Deliverables
● Develop a prototype ● Test functionality ● Submit provisional intellectual patent ● Submit abstract to national BMES
conferenceThis image cannot currently be displayed.
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Future Design Application
1 2 3
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http://www.nature.com/srep/2013/130521/srep01863/full/srep01863.html#supplementary-informationhttp://www.sciencedirect.com/science/article/pii/S1549963413005893
Timeline17
Budget
http://www.biocompare.com/pfu/110447/soids/19788/Antibodies/PBP2ahttp://www.hometrainingtools.com/checkout/cart/http://www.everbrightsolar.net/sylgard-184-silicone-elastomer-kit-encapsulation-184.htmlhttp://www.walmart.com/ip/Ranger-Pipette-Pop-Refill-Pack-25-pcs/26682149http://www.amazon.com/Jensen-Global-Syringe-Manual-Calibrated/dp/B002R18F02/ref=sr_1_8?ie=UTF8&qid=1418330917&sr=8-8&keywords=30+gauge+syringehttp://www.amazon.com/TOOGOO-Plastic-Stainless-Dispenser-0-176mm/dp/B00J2N3OK4/ref=sr_1_5?ie=UTF8&qid=1418331020&sr=8-5&keywords=30+gauge+needles#product-description-iframehttp://www.amazon.com/Latex-Tubing-I-D-O-D-WALL/dp/B0045V36PQhttp://www.abcam.com/pbp-antibody-ab58224.html
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Team Structure
Amanda Kaitlyn
OSHA Training
Fabricating
Fluid Dynamics
Antibody Immobilization
Testing
Device Design
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Acknowledgements I.)Wayne State University’s Smart Sensors and Integrated Microsystems (SSIM)
a.Dr. Auner II.)Lawrence Technological University
a.Dr. Li b.Dr. Nasir c. Dr. Bou- Akl
III.)The LEGENDS Entrepreneurial Student Awards
20
Questions?