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MRSA Diagnostics Kaitlyn Tingley & Amanda Bukhtia Lawrence Technological University Advisors: Dr. Li, Dr. Nasir, Dr. Bou-Akl and Dr. Auner

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Page 1: MRSA Diagnostics - Lawrence Technological University ... · PDF fileMRSA Diagnostics Kaitlyn Tingley & Amanda Bukhtia ... Adapted from Dr. Nasir Inlet Outlet PDMS Glass Au Electrodes

MRSA DiagnosticsKaitlyn Tingley & Amanda BukhtiaLawrence Technological University

Advisors: Dr. Li, Dr. Nasir, Dr. Bou-Akl and Dr. Auner

Page 2: MRSA Diagnostics - Lawrence Technological University ... · PDF fileMRSA Diagnostics Kaitlyn Tingley & Amanda Bukhtia ... Adapted from Dr. Nasir Inlet Outlet PDMS Glass Au Electrodes

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

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Page 3: MRSA Diagnostics - Lawrence Technological University ... · PDF fileMRSA Diagnostics Kaitlyn Tingley & Amanda Bukhtia ... Adapted from Dr. Nasir Inlet Outlet PDMS Glass Au Electrodes

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

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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

Page 8: MRSA Diagnostics - Lawrence Technological University ... · PDF fileMRSA Diagnostics Kaitlyn Tingley & Amanda Bukhtia ... Adapted from Dr. Nasir Inlet Outlet PDMS Glass Au Electrodes

Design Concept

.

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Page 9: MRSA Diagnostics - Lawrence Technological University ... · PDF fileMRSA Diagnostics Kaitlyn Tingley & Amanda Bukhtia ... Adapted from Dr. Nasir Inlet Outlet PDMS Glass Au Electrodes

Slide 8

1 Finish NotesKaitlyn Tingley,

Page 10: MRSA Diagnostics - Lawrence Technological University ... · PDF fileMRSA Diagnostics Kaitlyn Tingley & Amanda Bukhtia ... Adapted from Dr. Nasir Inlet Outlet PDMS Glass Au Electrodes

Project Tasks

1. Device Fabrication 2. Antibody Immobilization3. Simulation to determine flow parameters 4. Device Testing

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Slide 9

2 Finish NotesKaitlyn Tingley,

Page 12: MRSA Diagnostics - Lawrence Technological University ... · PDF fileMRSA Diagnostics Kaitlyn Tingley & Amanda Bukhtia ... Adapted from Dr. Nasir Inlet Outlet PDMS Glass Au Electrodes

Electrode Fabrication

• Substrate Material: Glass (slide)

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Slide 10

3 Finish NotesKaitlyn Tingley,

Page 14: MRSA Diagnostics - Lawrence Technological University ... · PDF fileMRSA Diagnostics Kaitlyn Tingley & Amanda Bukhtia ... Adapted from Dr. Nasir Inlet Outlet PDMS Glass Au Electrodes

Flow Channel Fabrication

● Soft Lithography

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Slide 11

4 Finish NotesKaitlyn Tingley,

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Antibody Immobilization and Device Bonding11

1. Antibody Immobili-zation 2. Device Bonding

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Device Testing12

Adapted from Dr. Nasir

Inlet Outlet

PDMS

Glass

Au Electrodes Non-MRSAMRSA

5

Page 18: MRSA Diagnostics - Lawrence Technological University ... · PDF fileMRSA Diagnostics Kaitlyn Tingley & Amanda Bukhtia ... Adapted from Dr. Nasir Inlet Outlet PDMS Glass Au Electrodes

Slide 13

5 Finish NotesKaitlyn Tingley,

Page 19: MRSA Diagnostics - Lawrence Technological University ... · PDF fileMRSA Diagnostics Kaitlyn Tingley & Amanda Bukhtia ... Adapted from Dr. Nasir Inlet Outlet PDMS Glass Au Electrodes

Testing Parameters

• Staph SolutionMSSA (Control)MRSA(Variable)

• Input: Staph Solution

• Output: Impedance

1367

Page 20: MRSA Diagnostics - Lawrence Technological University ... · PDF fileMRSA Diagnostics Kaitlyn Tingley & Amanda Bukhtia ... Adapted from Dr. Nasir Inlet Outlet PDMS Glass Au Electrodes

Slide 14

6 Finish NotesKaitlyn Tingley,

7 Double check last sentenceKaitlyn Tingley,

Page 21: MRSA Diagnostics - Lawrence Technological University ... · PDF fileMRSA Diagnostics Kaitlyn Tingley & Amanda Bukhtia ... Adapted from Dr. Nasir Inlet Outlet PDMS Glass Au Electrodes

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

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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

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Timeline17

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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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Page 27: MRSA Diagnostics - Lawrence Technological University ... · PDF fileMRSA Diagnostics Kaitlyn Tingley & Amanda Bukhtia ... Adapted from Dr. Nasir Inlet Outlet PDMS Glass Au Electrodes

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

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Questions?