Presentation biomedical virus detection_prabeen_kattel

Nano Particles for pathogen detection

Prabeen KattelBiomedical instrumentation and Electrical safety

Instructor: Dr. Hyeun Joong Yoon

Outline• Introduction • Methods and materials • Result and analysis• conclusion

Research articles• Graphene-based electrochemical biosensor for pathogenic

virus detection(Fei Liu, Ki Seok Choi, Tae Jung Park, Sang Yup Lee & Tae Seok Seo)• Graphene oxide based immunobiosensor for ultra sensitive

pathogen detection(Jae Hwan Jung,Fei Liu,and Tae Seok Seo)• Micropatterned reduced graphene oxide based field-effect

transistor for real-time virus detection(Fei Liu , Yo Han Kim , Doo Sung Cheon , Tae Seok Seo) • Real-Time Capture and Visualization of Individual Viruses in

Complex Media (Steven M. Scherr, George G. Daaboul, Jacob Trueb, Derin Sevenler, Helen Fawcett, Bennett Goldberg, John H. Connor, and M. Selim Unlu )

Nano particles

• Carbon based Particles:- Graphene and nanotubes• Metal Based Particles: - quantum dots,nanogold,nanosilver• Dendrimers: -Nanosized polymers built from branches units• Composites: - Combine one nanoparticles with another nano particles or larger , bulk type material.

Carbon Nano tube for trapping virus

Scanning electron microscope image (scale bar, 200 nm) of the H5N2 avian influenza virus (purple) trapped inside the aligned carbon nanotubes.

Credit: Image courtesy of Penn State University

https://images.sciencedaily.com/2016/10/161010143745_1_900x600.jpg

Graphene Preparation

(A) Fabrication of GO films by the centrifugal vacuum evaporation method.

(B) freestanding GO film (top), and the reduced GO film (bottom).

(C) SEM image of the reduced GO film

(D) A representative cyclic voltammograms by using the bare graphene film as an electrochemical electrode in the [Fe(CN)6]3-/4- redox system

https://www.researchgate.net/publication/226430993_Graphene based _electrochemical_ biosensor_for_pathogenic _virus_detection

https://www.researchgate.net/publication/226430993_Graphene%20based_electrochemical_biosensor_for_pathogenic_virus_detection




Fabrication with graphene electrodePSE modificationAntibody ImmobilizationVirus capture

https://www.researchgate.net/publication/226430993_Graphene based electrochemical_ biosensor_for_pathogenic _virus_detection

Graphene film-based biosensor for pathogen virus detection





Graphene oxide based immunobiosensor for ultra sensitive pathogen detection

• Synthesis of Ab-DNA-Au NPS conjugate• Immobilization of antibodies• Pathogen Detection by illuminance measurement.• Characterization of the GO surface using AFM images of modified GO array

http://ieeexplore.ieee.org.excelsior.sdstate.edu/document/5697935/

Micropatterned reduced graphene oxide based field-effect transistor for real-time virus detection

• Fabrication of a large area graphene oxide (LAGO)• Fabrication of an(micropatterned reduced graphene oxide field-effect

transistor ) MRGO-FET device

- http://www.sciencedirect.com/science/article/pii/S0925400513006801

http://www.sciencedirect.com/science/article/pii/S0925400513006801

Virus detection on the MRGO-FET

• specific rotavirus capture was monitored by observing the current changes • Five independent

experiments were performed for each concentration of virus samples ranging from 10 to 105 pfu/mL



Real-Time Capture and Visualization of Individual Viruses in Complex Media

• An imaging technique for real-time, sensitive, and label-free visualization of viruses and nanoparticles directly in complex solutions such as serum

• single-particle interferometric imaging sensing(SP-IRIS)

• Anti Ebola antibody capture probes (13F6) is printed on the surface of the sensor • capture and visualization of a recombinant vesicular stomatitis virus Ebola model (rVSV-ZEBOV) at 100 PFU/mL in

undiluted fetal bovine serum in less than 30 min

http://pubs.acs.org/doi/abs/10.1021/acsnano.5b07948


Real-Time Detection of Individual Viruses.

Using this microfluidic chamber, a solution of PBS containing virus was directed through the fluid channel

Images of the IRIS chip surface were recorded every 30 s. Following image acquisition, individual particles were counted using custom particle detection

software.http://pubs.acs.org/doi/abs/10.1021/acsnano.5b07948


Cyclic voltammogram of the bare graphene electrode with different combination






Anodic peak current change depending on the input cell

number of the rotavirus from 10 to 103 pfu






Quenching Effect at various concentration of rotavirus on immunosensor

http://ieeexplore.ieee.org.excelsior.sdstate.edu/document/5697935/

Result of MRGO-FET based biosensor for rotavirus detection


Real time and label free virus detection on MRGO-FET device Current response of device depending on the concentration of Rotavirus blank(black), 10(red), 10^2(blue), 10^3(green), 10^4(pink), 10^5 pfu/mL(yellow)


Virus Detection Sensitivity Directly in Buffer Solution by SP-IRIS

• determined the performance and specificity of the sensor system• conducted dilution experiments for rVSV-ZEBOV spiked in PBS with 1% BSA

and measured the limit of detection.• six different dilutions were made from the stock virus solution



Virus Detection and Quantification Directly in Complex Solutions

• capability and specificity of SPIRIS in direct label-free detection of viruses in 100% fetal bovine serum is measured again

• Track very closely with the dilution curve in PBS. • linear curve was fit to the time-resolved virus count for each concentration.

Figure: Accumulation of rVSV-ZEBOV virus for a serial dilution

Conclusion• Nano Materials like Au and Graphene are found to

be very effective in pathogen detection because of its electrical properties.• Graphene based system is used in detecting

Rotavirus and Copmlex solution based detection is used for virus Ebola model (rVSV-ZEBOV).• Graphene based detection can utilize either

reduced current measurement techniques or quenching effect measurement.

Science

Presentation biomedical virus detection_prabeen_kattel