Team 19 Kaitlyn Clarke Brittany DePoi Andrew Reynolds Adarsha Selvachandran Client: Dr. Wei Sun.
Amanda Bukhtia and Kaitlyn Tingley Lawrence · PDF fileAmanda Bukhtia and Kaitlyn Tingley...
Transcript of Amanda Bukhtia and Kaitlyn Tingley Lawrence · PDF fileAmanda Bukhtia and Kaitlyn Tingley...
![Page 1: Amanda Bukhtia and Kaitlyn Tingley Lawrence · PDF fileAmanda Bukhtia and Kaitlyn Tingley Lawrence Technological University Advisors: Dr. Auner, Dr. Bou-Akl, Dr. Li and Dr. Nasir.](https://reader033.fdocuments.us/reader033/viewer/2022051722/5aa9e8e27f8b9a81188d5408/html5/thumbnails/1.jpg)
Amanda Bukhtia and Kaitlyn Tingley
Lawrence Technological University
Advisors: Dr. Auner, Dr. Bou-Akl, Dr. Li and Dr. Nasir
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Need
“Develop a quick and cost effective method to detect if a
patient has (MRSA)”
Solution
• A novel impedance biosensor
Project Summary
.
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What is MRSA?
An antibiotic resistant
staph infection
Can enter the bloodstream
through open wounds
which can lead to death
75,309 people are
estimated to have MRSA
every year
http://www.cdc.gov/mrsa/tracking/index.html
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Current Diagnostic Methods3
http://laboratory-manager.advanceweb.com/Archives/Article-Archives/MRSA-Screening-Culture-or-Molecular.aspx
http://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/1471-2180/6/83
https://workspace.imperial.ac.uk/opticalandsemidev/public/Publications/LAWES.pdf
http://www.aist.go.jp/aist_e/latest_research/2012/20121121/20121121.html
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• Penicillin binding protein
2A (PBP2a)
• Responsible to antibiotic
resistance in MRSA
• Regulates the cell wall of
MRSA bacteria
PBP-2a Antibody4
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Device Design5
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Device Design
.
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Device A Device B
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Device Fabrication
Glass Wafer Fabrication1. Electrodes (Au)
2. Passivation (SiO2)
PDMS Fabrication
Device Bonding
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Device A Device B
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Electrode Fabrication8
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Passivation Layer Fabrication9
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Channel Fabrication10
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Device Bonding
• Oxygen plasma
treated PDMS and
glass
• Bonded through
minimal compression
and heat
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Impedance Testing12
Impedance
Analyzer
Syringe
Pump
Device
Probe
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Impedance Testing (Part 1)
• Four different concentrations of PBS
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Impedance Testing (Part 2)
• Impedance (Ω) is inversely hyperbolic to ion concentration
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Impedance Testing (Part 3)
• Device was placed on
magnetic platform
• Steel particles were
mixed into a 1x PBS
solution
• Impedance decreases
as steel particles are
captivated by the magnet
onto the electrode
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Antibody Binding Testing
Testing Conditions
• Three concentrations of anti-PBP-2a
• Whole MRSA
- Experimental
• Whole methicillin-sensitive staphylococcus aureus
MSSA
-Control
• Fluorescent staining of antibody/bacteria complex
-Using Alexa Fluor
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Antibody Binding Testing17
Results
• Positive detection of anti-PBP-2a/<RSA complex
-Green fluorescence
• Failure to detect anti-PBP-2a/MSSA
-No fluorescence
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Antibody Immobilization 18
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Results19
• PBP-2a antibody will bind to MRSA bacteria but not to
MSSA
- This shows the specificity of our biological reagent
• Fluid dynamics and impedance mechanisms of device
have been validated
- Device A provides superior results due to enhanced
PDMS adhesion
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Timeline20
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Team Structure
Amanda Kaitlyn
OSHA Training
Fabrication
Impedance
Testing
Antibody
Immobilization
Antibody
Binding Testing
Device Design
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Future Outlook22
http://www.nature.com/srep/2013/130521/srep01863/full/srep01863.html#suppl
ementary-information
http://www.sciencedirect.com/science/article/pii/S1549963413005893
Establish flow through testing conditions using
COMSOL modeling
Validate anti-PBP-2a immobilization
Conduct device impedance testing with MRSA
Integrate impedance analyzer
Create packaging for device
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Acknowledgements
Wayne State University’s Smart Sensors and Integrated
Microsystems (SSIM)
Dr. Auner
Lawrence Technological University
Dr. Li
Dr. Nasir
Dr. Bou-akl
The LEGENDS Entrepreneurial Student Awards
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Questions?
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MRSA Diagnostics Amanda Bukhtia and Kaitlyn TingleyDr. Auner, Dr. Bou-Akl, Dr. Li and Dr. Nasir
Objective: To create a novel device for the rapid detection of MRSA.
Approach: Create a microfluidic device to detect MRSA due to these devices quick turn around time and their cheap manufacturing costs.
Impact: Began proof of concept for a novel solution to detect MRSA quickly while maintaining high specificity
Progress and Results:
1. We have validated the impedance mechanism of our device
2. We have verified the specificity mechanism of our device
3. We have shown device A has better binding properties
Antibody Testing
Impedance Testing