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2021
Zhang, Yi; Yurdakul, Celalettin; Devaux, Alexander J; Wang, Le; Xu, Xiaoji G; Connor, John H; Ünlü, M Selim; Cheng, Ji-Xin
Vibrational spectroscopic detection of a single virus by mid-infrared photothermal microscopy Journal Article
In: Analytical Chemistry, vol. 92, iss. 8, pp. 4100, 2021.
Abstract | Links | BibTeX | Tags: Virus
@article{nokey,
title = {Vibrational spectroscopic detection of a single virus by mid-infrared photothermal microscopy},
author = {Yi Zhang and Celalettin Yurdakul and Alexander J Devaux and Le Wang and Xiaoji G Xu and John H Connor and M Selim Ünlü and Ji-Xin Cheng},
url = {https://sites.bu.edu/cheng-group/files/2021/02/277-analytical-chemistry-single-virus-detection-by-MIP.pdf},
doi = {https://dx.doi.org/10.1021/acs.analchem.0c05333},
year = {2021},
date = {2021-02-17},
urldate = {2021-02-17},
journal = {Analytical Chemistry},
volume = {92},
issue = {8},
pages = {4100},
abstract = {We report a confocal interferometric mid-infrared photothermal (MIP) microscope for ultra-sensitive and spatially resolved chemical imaging of individual viruses. The interferometric scattering principle is applied to detect the very weak photothermal signal induced by infrared absorption of chemical bonds. Spectroscopic MIP detection of single vesicular stomatitis viruses (VSVs) and poxviruses is demonstrated. The single virus spectra show high consistency within the same virus type. The dominant spectral peaks are contributed by the amide I and amide II vibrations attributed to the viral proteins. The ratio of these two peaks is significantly different between VSVs and poxviruses, highlighting the potential of using interferometric MIP microscopy for label-free differentiation of viral particles. This all-optical chemical imaging method opens a new way for spectroscopic detection of biological nanoparticles in a label-free manner and may facilitate in predicting and controlling the outbreaks of emerging virus strains.},
keywords = {Virus},
pubstate = {published},
tppubtype = {article}
}
We report a confocal interferometric mid-infrared photothermal (MIP) microscope for ultra-sensitive and spatially resolved chemical imaging of individual viruses. The interferometric scattering principle is applied to detect the very weak photothermal signal induced by infrared absorption of chemical bonds. Spectroscopic MIP detection of single vesicular stomatitis viruses (VSVs) and poxviruses is demonstrated. The single virus spectra show high consistency within the same virus type. The dominant spectral peaks are contributed by the amide I and amide II vibrations attributed to the viral proteins. The ratio of these two peaks is significantly different between VSVs and poxviruses, highlighting the potential of using interferometric MIP microscopy for label-free differentiation of viral particles. This all-optical chemical imaging method opens a new way for spectroscopic detection of biological nanoparticles in a label-free manner and may facilitate in predicting and controlling the outbreaks of emerging virus strains.
Zhang, Yi; Yurdakul, Celalettin; Devaux, Alexander J; Wang, Le; Xu, Xiaoji G; Connor, John H; Ünlü, M Selim; Cheng, Ji-Xin
Vibrational spectroscopic detection of a single virus by mid-infrared photothermal microscopy Journal Article
In: Analytical Chemistry, vol. 92, iss. 8, pp. 4100, 2021.
Abstract | Links | BibTeX | Tags: Virus
@article{nokeyl,
title = {Vibrational spectroscopic detection of a single virus by mid-infrared photothermal microscopy},
author = {Yi Zhang and Celalettin Yurdakul and Alexander J Devaux and Le Wang and Xiaoji G Xu and John H Connor and M Selim Ünlü and Ji-Xin Cheng},
url = {https://sites.bu.edu/cheng-group/files/2021/02/277-analytical-chemistry-single-virus-detection-by-MIP.pdf},
doi = {https://dx.doi.org/10.1021/acs.analchem.0c05333},
year = {2021},
date = {2021-02-17},
urldate = {2021-02-17},
journal = {Analytical Chemistry},
volume = {92},
issue = {8},
pages = {4100},
abstract = {We report a confocal interferometric mid-infrared photothermal (MIP) microscope for ultra-sensitive and spatially resolved chemical imaging of individual viruses. The interferometric scattering principle is applied to detect the very weak photothermal signal induced by infrared absorption of chemical bonds. Spectroscopic MIP detection of single vesicular stomatitis viruses (VSVs) and poxviruses is demonstrated. The single virus spectra show high consistency within the same virus type. The dominant spectral peaks are contributed by the amide I and amide II vibrations attributed to the viral proteins. The ratio of these two peaks is significantly different between VSVs and poxviruses, highlighting the potential of using interferometric MIP microscopy for label-free differentiation of viral particles. This all-optical chemical imaging method opens a new way for spectroscopic detection of biological nanoparticles in a label-free manner and may facilitate in predicting and controlling the outbreaks of emerging virus strains.},
keywords = {Virus},
pubstate = {published},
tppubtype = {article}
}
We report a confocal interferometric mid-infrared photothermal (MIP) microscope for ultra-sensitive and spatially resolved chemical imaging of individual viruses. The interferometric scattering principle is applied to detect the very weak photothermal signal induced by infrared absorption of chemical bonds. Spectroscopic MIP detection of single vesicular stomatitis viruses (VSVs) and poxviruses is demonstrated. The single virus spectra show high consistency within the same virus type. The dominant spectral peaks are contributed by the amide I and amide II vibrations attributed to the viral proteins. The ratio of these two peaks is significantly different between VSVs and poxviruses, highlighting the potential of using interferometric MIP microscopy for label-free differentiation of viral particles. This all-optical chemical imaging method opens a new way for spectroscopic detection of biological nanoparticles in a label-free manner and may facilitate in predicting and controlling the outbreaks of emerging virus strains.
Seymour, Elif; Ünlü, Nese Lortlar; Carter, Erik P; Connor, John H; Ünlü, M Selim
Configurable Digital Virus Counter on Robust Universal DNA Chips Journal Article
In: ACS sensors, vol. 6, iss. 10, pp. 229, 2021.
Abstract | Links | BibTeX | Tags: Virus
@article{nokey,
title = {Configurable Digital Virus Counter on Robust Universal DNA Chips},
author = {Elif Seymour and Nese Lortlar Ünlü and Erik P Carter and John H Connor and M Selim Ünlü},
url = {https://www.biorxiv.org/content/biorxiv/early/2020/10/22/2020.10.22.350579.full.pdf},
doi = {https://doi.org/10.1101/2020.10.22.350579},
year = {2021},
date = {2021-01-11},
urldate = {2021-01-11},
journal = {ACS sensors},
volume = {6},
issue = {10},
pages = {229},
abstract = {Here, we demonstrate real-time multiplexed virus detection by applying DNA-directed antibody immobilization technique to a single-particle interferometric reflectance imaging sensor (SP-IRIS). In this technique, the biosensor chip surface spotted with different DNA sequences is converted to a multiplexed antibody array by flowing antibody-DNA conjugates and allowing specific DNA-DNA hybridization. The resulting antibody array is shown to detect three different recombinant Vesicular Stomatitis Viruses (rVSVs) genetically engineered to express surface glycoproteins of Ebola, Marburg, and Lassa viruses in real-time in a disposable microfluidic cartridge. We also show that this method can be modified to produce a single-step, homogeneous assay format by mixing the antibody-DNA conjugates with the virus sample in solution phase prior to flowing in the microfluidic cartridge, eliminating the antibody immobilization step. This homogenous approach achieved detection of the model Ebola virus, rVSV-EBOV, at a concentration of 100 PFU/ml in 1 hour. Finally, we demonstrate the feasibility of this homogeneous technique as a rapid test using a passive microfluidic cartridge. A concentration of 104 PFU/ml was detectable under 10 minutes for the rVSV-Ebola virus. Utilizing DNA microarrays for antibody-based diagnostics is an alternative approach to antibody microarrays and offers advantages such as configurable sensor surface, long-term storage ability, and decreased antibody use. We believe these properties will make SP-IRIS a versatile and robust platform for point-of-care diagnostics applications.
},
keywords = {Virus},
pubstate = {published},
tppubtype = {article}
}
Here, we demonstrate real-time multiplexed virus detection by applying DNA-directed antibody immobilization technique to a single-particle interferometric reflectance imaging sensor (SP-IRIS). In this technique, the biosensor chip surface spotted with different DNA sequences is converted to a multiplexed antibody array by flowing antibody-DNA conjugates and allowing specific DNA-DNA hybridization. The resulting antibody array is shown to detect three different recombinant Vesicular Stomatitis Viruses (rVSVs) genetically engineered to express surface glycoproteins of Ebola, Marburg, and Lassa viruses in real-time in a disposable microfluidic cartridge. We also show that this method can be modified to produce a single-step, homogeneous assay format by mixing the antibody-DNA conjugates with the virus sample in solution phase prior to flowing in the microfluidic cartridge, eliminating the antibody immobilization step. This homogenous approach achieved detection of the model Ebola virus, rVSV-EBOV, at a concentration of 100 PFU/ml in 1 hour. Finally, we demonstrate the feasibility of this homogeneous technique as a rapid test using a passive microfluidic cartridge. A concentration of 104 PFU/ml was detectable under 10 minutes for the rVSV-Ebola virus. Utilizing DNA microarrays for antibody-based diagnostics is an alternative approach to antibody microarrays and offers advantages such as configurable sensor surface, long-term storage ability, and decreased antibody use. We believe these properties will make SP-IRIS a versatile and robust platform for point-of-care diagnostics applications.
Seymour, Elif; Ünlü, Nese Lortlar; Carter, Erik P; Connor, John H; Ünlü, M Selim
Configurable Digital Virus Counter on Robust Universal DNA Chips Journal Article
In: ACS sensors, vol. 6, iss. 10, pp. 229, 2021.
Abstract | Links | BibTeX | Tags: Virus
@article{nokeym,
title = {Configurable Digital Virus Counter on Robust Universal DNA Chips},
author = {Elif Seymour and Nese Lortlar Ünlü and Erik P Carter and John H Connor and M Selim Ünlü},
url = {https://www.biorxiv.org/content/biorxiv/early/2020/10/22/2020.10.22.350579.full.pdf},
doi = {https://doi.org/10.1101/2020.10.22.350579},
year = {2021},
date = {2021-01-11},
urldate = {2021-01-11},
journal = {ACS sensors},
volume = {6},
issue = {10},
pages = {229},
abstract = {Here, we demonstrate real-time multiplexed virus detection by applying DNA-directed antibody immobilization technique to a single-particle interferometric reflectance imaging sensor (SP-IRIS). In this technique, the biosensor chip surface spotted with different DNA sequences is converted to a multiplexed antibody array by flowing antibody-DNA conjugates and allowing specific DNA-DNA hybridization. The resulting antibody array is shown to detect three different recombinant Vesicular Stomatitis Viruses (rVSVs) genetically engineered to express surface glycoproteins of Ebola, Marburg, and Lassa viruses in real-time in a disposable microfluidic cartridge. We also show that this method can be modified to produce a single-step, homogeneous assay format by mixing the antibody-DNA conjugates with the virus sample in solution phase prior to flowing in the microfluidic cartridge, eliminating the antibody immobilization step. This homogenous approach achieved detection of the model Ebola virus, rVSV-EBOV, at a concentration of 100 PFU/ml in 1 hour. Finally, we demonstrate the feasibility of this homogeneous technique as a rapid test using a passive microfluidic cartridge. A concentration of 104 PFU/ml was detectable under 10 minutes for the rVSV-Ebola virus. Utilizing DNA microarrays for antibody-based diagnostics is an alternative approach to antibody microarrays and offers advantages such as configurable sensor surface, long-term storage ability, and decreased antibody use. We believe these properties will make SP-IRIS a versatile and robust platform for point-of-care diagnostics applications.},
keywords = {Virus},
pubstate = {published},
tppubtype = {article}
}
Here, we demonstrate real-time multiplexed virus detection by applying DNA-directed antibody immobilization technique to a single-particle interferometric reflectance imaging sensor (SP-IRIS). In this technique, the biosensor chip surface spotted with different DNA sequences is converted to a multiplexed antibody array by flowing antibody-DNA conjugates and allowing specific DNA-DNA hybridization. The resulting antibody array is shown to detect three different recombinant Vesicular Stomatitis Viruses (rVSVs) genetically engineered to express surface glycoproteins of Ebola, Marburg, and Lassa viruses in real-time in a disposable microfluidic cartridge. We also show that this method can be modified to produce a single-step, homogeneous assay format by mixing the antibody-DNA conjugates with the virus sample in solution phase prior to flowing in the microfluidic cartridge, eliminating the antibody immobilization step. This homogenous approach achieved detection of the model Ebola virus, rVSV-EBOV, at a concentration of 100 PFU/ml in 1 hour. Finally, we demonstrate the feasibility of this homogeneous technique as a rapid test using a passive microfluidic cartridge. A concentration of 104 PFU/ml was detectable under 10 minutes for the rVSV-Ebola virus. Utilizing DNA microarrays for antibody-based diagnostics is an alternative approach to antibody microarrays and offers advantages such as configurable sensor surface, long-term storage ability, and decreased antibody use. We believe these properties will make SP-IRIS a versatile and robust platform for point-of-care diagnostics applications.
1.
Zhang, Yi; Yurdakul, Celalettin; Devaux, Alexander J; Wang, Le; Xu, Xiaoji G; Connor, John H; Ünlü, M Selim; Cheng, Ji-Xin
Vibrational spectroscopic detection of a single virus by mid-infrared photothermal microscopy Journal Article
In: Analytical Chemistry, vol. 92, iss. 8, pp. 4100, 2021.
Abstract | Links | BibTeX | Tags: Virus
@article{nokey,
title = {Vibrational spectroscopic detection of a single virus by mid-infrared photothermal microscopy},
author = {Yi Zhang and Celalettin Yurdakul and Alexander J Devaux and Le Wang and Xiaoji G Xu and John H Connor and M Selim Ünlü and Ji-Xin Cheng},
url = {https://sites.bu.edu/cheng-group/files/2021/02/277-analytical-chemistry-single-virus-detection-by-MIP.pdf},
doi = {https://dx.doi.org/10.1021/acs.analchem.0c05333},
year = {2021},
date = {2021-02-17},
urldate = {2021-02-17},
journal = {Analytical Chemistry},
volume = {92},
issue = {8},
pages = {4100},
abstract = {We report a confocal interferometric mid-infrared photothermal (MIP) microscope for ultra-sensitive and spatially resolved chemical imaging of individual viruses. The interferometric scattering principle is applied to detect the very weak photothermal signal induced by infrared absorption of chemical bonds. Spectroscopic MIP detection of single vesicular stomatitis viruses (VSVs) and poxviruses is demonstrated. The single virus spectra show high consistency within the same virus type. The dominant spectral peaks are contributed by the amide I and amide II vibrations attributed to the viral proteins. The ratio of these two peaks is significantly different between VSVs and poxviruses, highlighting the potential of using interferometric MIP microscopy for label-free differentiation of viral particles. This all-optical chemical imaging method opens a new way for spectroscopic detection of biological nanoparticles in a label-free manner and may facilitate in predicting and controlling the outbreaks of emerging virus strains.},
keywords = {Virus},
pubstate = {published},
tppubtype = {article}
}
We report a confocal interferometric mid-infrared photothermal (MIP) microscope for ultra-sensitive and spatially resolved chemical imaging of individual viruses. The interferometric scattering principle is applied to detect the very weak photothermal signal induced by infrared absorption of chemical bonds. Spectroscopic MIP detection of single vesicular stomatitis viruses (VSVs) and poxviruses is demonstrated. The single virus spectra show high consistency within the same virus type. The dominant spectral peaks are contributed by the amide I and amide II vibrations attributed to the viral proteins. The ratio of these two peaks is significantly different between VSVs and poxviruses, highlighting the potential of using interferometric MIP microscopy for label-free differentiation of viral particles. This all-optical chemical imaging method opens a new way for spectroscopic detection of biological nanoparticles in a label-free manner and may facilitate in predicting and controlling the outbreaks of emerging virus strains.
2.
Zhang, Yi; Yurdakul, Celalettin; Devaux, Alexander J; Wang, Le; Xu, Xiaoji G; Connor, John H; Ünlü, M Selim; Cheng, Ji-Xin
Vibrational spectroscopic detection of a single virus by mid-infrared photothermal microscopy Journal Article
In: Analytical Chemistry, vol. 92, iss. 8, pp. 4100, 2021.
Abstract | Links | BibTeX | Tags: Virus
@article{nokeyl,
title = {Vibrational spectroscopic detection of a single virus by mid-infrared photothermal microscopy},
author = {Yi Zhang and Celalettin Yurdakul and Alexander J Devaux and Le Wang and Xiaoji G Xu and John H Connor and M Selim Ünlü and Ji-Xin Cheng},
url = {https://sites.bu.edu/cheng-group/files/2021/02/277-analytical-chemistry-single-virus-detection-by-MIP.pdf},
doi = {https://dx.doi.org/10.1021/acs.analchem.0c05333},
year = {2021},
date = {2021-02-17},
urldate = {2021-02-17},
journal = {Analytical Chemistry},
volume = {92},
issue = {8},
pages = {4100},
abstract = {We report a confocal interferometric mid-infrared photothermal (MIP) microscope for ultra-sensitive and spatially resolved chemical imaging of individual viruses. The interferometric scattering principle is applied to detect the very weak photothermal signal induced by infrared absorption of chemical bonds. Spectroscopic MIP detection of single vesicular stomatitis viruses (VSVs) and poxviruses is demonstrated. The single virus spectra show high consistency within the same virus type. The dominant spectral peaks are contributed by the amide I and amide II vibrations attributed to the viral proteins. The ratio of these two peaks is significantly different between VSVs and poxviruses, highlighting the potential of using interferometric MIP microscopy for label-free differentiation of viral particles. This all-optical chemical imaging method opens a new way for spectroscopic detection of biological nanoparticles in a label-free manner and may facilitate in predicting and controlling the outbreaks of emerging virus strains.},
keywords = {Virus},
pubstate = {published},
tppubtype = {article}
}
We report a confocal interferometric mid-infrared photothermal (MIP) microscope for ultra-sensitive and spatially resolved chemical imaging of individual viruses. The interferometric scattering principle is applied to detect the very weak photothermal signal induced by infrared absorption of chemical bonds. Spectroscopic MIP detection of single vesicular stomatitis viruses (VSVs) and poxviruses is demonstrated. The single virus spectra show high consistency within the same virus type. The dominant spectral peaks are contributed by the amide I and amide II vibrations attributed to the viral proteins. The ratio of these two peaks is significantly different between VSVs and poxviruses, highlighting the potential of using interferometric MIP microscopy for label-free differentiation of viral particles. This all-optical chemical imaging method opens a new way for spectroscopic detection of biological nanoparticles in a label-free manner and may facilitate in predicting and controlling the outbreaks of emerging virus strains.
3.
Seymour, Elif; Ünlü, Nese Lortlar; Carter, Erik P; Connor, John H; Ünlü, M Selim
Configurable Digital Virus Counter on Robust Universal DNA Chips Journal Article
In: ACS sensors, vol. 6, iss. 10, pp. 229, 2021.
Abstract | Links | BibTeX | Tags: Virus
@article{nokey,
title = {Configurable Digital Virus Counter on Robust Universal DNA Chips},
author = {Elif Seymour and Nese Lortlar Ünlü and Erik P Carter and John H Connor and M Selim Ünlü},
url = {https://www.biorxiv.org/content/biorxiv/early/2020/10/22/2020.10.22.350579.full.pdf},
doi = {https://doi.org/10.1101/2020.10.22.350579},
year = {2021},
date = {2021-01-11},
urldate = {2021-01-11},
journal = {ACS sensors},
volume = {6},
issue = {10},
pages = {229},
abstract = {Here, we demonstrate real-time multiplexed virus detection by applying DNA-directed antibody immobilization technique to a single-particle interferometric reflectance imaging sensor (SP-IRIS). In this technique, the biosensor chip surface spotted with different DNA sequences is converted to a multiplexed antibody array by flowing antibody-DNA conjugates and allowing specific DNA-DNA hybridization. The resulting antibody array is shown to detect three different recombinant Vesicular Stomatitis Viruses (rVSVs) genetically engineered to express surface glycoproteins of Ebola, Marburg, and Lassa viruses in real-time in a disposable microfluidic cartridge. We also show that this method can be modified to produce a single-step, homogeneous assay format by mixing the antibody-DNA conjugates with the virus sample in solution phase prior to flowing in the microfluidic cartridge, eliminating the antibody immobilization step. This homogenous approach achieved detection of the model Ebola virus, rVSV-EBOV, at a concentration of 100 PFU/ml in 1 hour. Finally, we demonstrate the feasibility of this homogeneous technique as a rapid test using a passive microfluidic cartridge. A concentration of 104 PFU/ml was detectable under 10 minutes for the rVSV-Ebola virus. Utilizing DNA microarrays for antibody-based diagnostics is an alternative approach to antibody microarrays and offers advantages such as configurable sensor surface, long-term storage ability, and decreased antibody use. We believe these properties will make SP-IRIS a versatile and robust platform for point-of-care diagnostics applications.
},
keywords = {Virus},
pubstate = {published},
tppubtype = {article}
}
Here, we demonstrate real-time multiplexed virus detection by applying DNA-directed antibody immobilization technique to a single-particle interferometric reflectance imaging sensor (SP-IRIS). In this technique, the biosensor chip surface spotted with different DNA sequences is converted to a multiplexed antibody array by flowing antibody-DNA conjugates and allowing specific DNA-DNA hybridization. The resulting antibody array is shown to detect three different recombinant Vesicular Stomatitis Viruses (rVSVs) genetically engineered to express surface glycoproteins of Ebola, Marburg, and Lassa viruses in real-time in a disposable microfluidic cartridge. We also show that this method can be modified to produce a single-step, homogeneous assay format by mixing the antibody-DNA conjugates with the virus sample in solution phase prior to flowing in the microfluidic cartridge, eliminating the antibody immobilization step. This homogenous approach achieved detection of the model Ebola virus, rVSV-EBOV, at a concentration of 100 PFU/ml in 1 hour. Finally, we demonstrate the feasibility of this homogeneous technique as a rapid test using a passive microfluidic cartridge. A concentration of 104 PFU/ml was detectable under 10 minutes for the rVSV-Ebola virus. Utilizing DNA microarrays for antibody-based diagnostics is an alternative approach to antibody microarrays and offers advantages such as configurable sensor surface, long-term storage ability, and decreased antibody use. We believe these properties will make SP-IRIS a versatile and robust platform for point-of-care diagnostics applications.
4.
Seymour, Elif; Ünlü, Nese Lortlar; Carter, Erik P; Connor, John H; Ünlü, M Selim
Configurable Digital Virus Counter on Robust Universal DNA Chips Journal Article
In: ACS sensors, vol. 6, iss. 10, pp. 229, 2021.
Abstract | Links | BibTeX | Tags: Virus
@article{nokeym,
title = {Configurable Digital Virus Counter on Robust Universal DNA Chips},
author = {Elif Seymour and Nese Lortlar Ünlü and Erik P Carter and John H Connor and M Selim Ünlü},
url = {https://www.biorxiv.org/content/biorxiv/early/2020/10/22/2020.10.22.350579.full.pdf},
doi = {https://doi.org/10.1101/2020.10.22.350579},
year = {2021},
date = {2021-01-11},
urldate = {2021-01-11},
journal = {ACS sensors},
volume = {6},
issue = {10},
pages = {229},
abstract = {Here, we demonstrate real-time multiplexed virus detection by applying DNA-directed antibody immobilization technique to a single-particle interferometric reflectance imaging sensor (SP-IRIS). In this technique, the biosensor chip surface spotted with different DNA sequences is converted to a multiplexed antibody array by flowing antibody-DNA conjugates and allowing specific DNA-DNA hybridization. The resulting antibody array is shown to detect three different recombinant Vesicular Stomatitis Viruses (rVSVs) genetically engineered to express surface glycoproteins of Ebola, Marburg, and Lassa viruses in real-time in a disposable microfluidic cartridge. We also show that this method can be modified to produce a single-step, homogeneous assay format by mixing the antibody-DNA conjugates with the virus sample in solution phase prior to flowing in the microfluidic cartridge, eliminating the antibody immobilization step. This homogenous approach achieved detection of the model Ebola virus, rVSV-EBOV, at a concentration of 100 PFU/ml in 1 hour. Finally, we demonstrate the feasibility of this homogeneous technique as a rapid test using a passive microfluidic cartridge. A concentration of 104 PFU/ml was detectable under 10 minutes for the rVSV-Ebola virus. Utilizing DNA microarrays for antibody-based diagnostics is an alternative approach to antibody microarrays and offers advantages such as configurable sensor surface, long-term storage ability, and decreased antibody use. We believe these properties will make SP-IRIS a versatile and robust platform for point-of-care diagnostics applications.},
keywords = {Virus},
pubstate = {published},
tppubtype = {article}
}
Here, we demonstrate real-time multiplexed virus detection by applying DNA-directed antibody immobilization technique to a single-particle interferometric reflectance imaging sensor (SP-IRIS). In this technique, the biosensor chip surface spotted with different DNA sequences is converted to a multiplexed antibody array by flowing antibody-DNA conjugates and allowing specific DNA-DNA hybridization. The resulting antibody array is shown to detect three different recombinant Vesicular Stomatitis Viruses (rVSVs) genetically engineered to express surface glycoproteins of Ebola, Marburg, and Lassa viruses in real-time in a disposable microfluidic cartridge. We also show that this method can be modified to produce a single-step, homogeneous assay format by mixing the antibody-DNA conjugates with the virus sample in solution phase prior to flowing in the microfluidic cartridge, eliminating the antibody immobilization step. This homogenous approach achieved detection of the model Ebola virus, rVSV-EBOV, at a concentration of 100 PFU/ml in 1 hour. Finally, we demonstrate the feasibility of this homogeneous technique as a rapid test using a passive microfluidic cartridge. A concentration of 104 PFU/ml was detectable under 10 minutes for the rVSV-Ebola virus. Utilizing DNA microarrays for antibody-based diagnostics is an alternative approach to antibody microarrays and offers advantages such as configurable sensor surface, long-term storage ability, and decreased antibody use. We believe these properties will make SP-IRIS a versatile and robust platform for point-of-care diagnostics applications.
