i mmunoassay utilizing magnetic beads for rapidvirus detection in the microfluidic flow cytometer...
TRANSCRIPT
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IMMUNOASSAY UTILIZING MAGNETIC BEADS FOR RAPIDVIRUS DETECTION IN THE MICROFLUIDIC FLOW CYTOMETER SYSTEM
Sung-Yi Yang, Kang-Yi Lien, Kao-Jean Huang, Huan-Yao Lei, Gwo-Bin Lee
National Cheng Kung University, Tainan Taiwan
uTAS2007
Professor : Liu, Cheng-Hsien Student : Olof Sterner
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AGENDA
1. Introduction2. Design and Fabrication3. Results4. Conclusion5. Discussion
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In this article: Integrated fluid chip for rapid
virus identification. Fluid movement is performed by
micro circulatory pump driven by airpressure.
Magnetic beads covered with specific antibodies.
Detection of virus is made through fluorescence by a PMT module.
INTRODUCTION
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Current viruses threatening mankind Influenza Hepatitis HIV Dengue fever Ebola Yellow Fever Etc
Treatment for viral infections are far behind those of bacterial infections Quick diagnosis crucial
INTRODUCTION
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DESIGN AND FABRICATION
Magnetic beads Covered in specific
antibodies toward a certain virus
Virus particle functions as antigen and binds specifically to the primary antibody on the magnetic bead.
Secondary antibody towards the same virus but targeting a different epitope binds to the virus and carries a fluorescent compound.
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DESIGN AND FABRICATION
Main detection principle1. Magnetic beads with
primary target antibody
2. Target antigen is added (Virus)
3. Secondary fluorescent antibodies is added
4. Fluidic focusing
5. Detection by means of fluorescense
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DESIGN AND FABRICATION
The integrated flow cytometer system
A
B
C
Dimensions : 46 mm * 66 mm * 5 mm
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DESIGN AND FABRICATION
A – Virus attachement to magnetic beads.
1. Injection of bio sample and magnetic beads
2. Introduction of sec fluorescent antibodies
3. Mixing with rotary microfluidic pump
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DESIGN AND FABRICATION
Airpressure driven rotary microfluidic pump
Biomed Microdevices (2007) 9:545–554DOI 10.1007/s10544-007-9062-6
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DESIGN AND FABRICATION
B – Washing and Pretreatment
1. Cu microcoils induced magnetic field attracts magnetic beads
2. Washing with buffer3. Release of magnetic beads(Flow generated by rotary microfluidic pump)
Purification and enrichment of virus samples utilizing magnetic beads on a microfluidic system, Lab Chip, DOI: 10.1039/b700516d, (2007)
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DESIGN AND FABRICATION
C – Fluid focusing, detection and sorting
1. Sheath flow injection2. Flow from rotary microfluidic pump for fluid
focusing3. Detection by PMT module4. Sorting of magnetic beads using feedback
signals from PMT5. Collection of magnetic beads(Flow generated by rotary microfluidic pump
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RESULTS
Evaluation of:
Pumping rate vs driving frequency of circulatory pump
Magnetic field and Temperatur vs Applied current
Fluorescence signal amplitude vs timeVirus detection limit
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RESULTS
Pumping rate vs driving frequence of circulatory pump
Magnetic field and Temperature in focusing vs Applied current
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RESULTS
Signal amplitude from fluorescence measurement vs time
Minimum concentration of 104 pfu/ml of Dengue virus could be detected.
RT-PCR has detection limit of ~1 pfu/ml
Singal to Noise ratio of 30
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CONCLUSION
Design for rapid virus
diagnosis has been proposed
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DISCUSSION
Easy principle, sandwich immunoassay is well documented
Gate design is not included in article
Sensitivity needs to be enchanced
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REFERENCES
Lab Chip, 2007, 7, 868–875, DOI: 10.1039/b700516d
Biomed Microdevices (2007) 9:545–554 DOI 10.1007/s10544-007-9062-6
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TACK!