tetano
Editor, Senior Moderator
Proc Natl Acad Sci U S A. 2019 Dec 27. pii: 201910113. doi: 10.1073/pnas.1910113117. [Epub ahead of print] [h=1]A rapid and label-free platform for virus capture and identification from clinical samples.[/h]
Yeh YT[SUP]1[/SUP], Gulino K[SUP]2[/SUP], Zhang Y[SUP]3[/SUP], Sabestien A[SUP]4[/SUP], Chou TW[SUP]2[/SUP], Zhou B[SUP]2[/SUP], Lin Z[SUP]3[/SUP], Albert I[SUP]4[/SUP], Lu H[SUP]5[/SUP], Swaminathan V[SUP]3[/SUP], Ghedin E[SUP]2[/SUP], Terrones M[SUP]1[/SUP].
[h=3]Author information[/h] 1 Department of Physics, The Pennsylvania State University, University Park, PA 16802; yxy155@psu.edu mut11@psu.edu. 2 Department of Biology, New York University, New York, NY 10003. 3 Department of Physics, The Pennsylvania State University, University Park, PA 16802. 4 Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA 16802. 5 Department of Veterinary and Biomedical Sciences, The Pennsylvania State University, University Park, PA 16802.
[h=3]Abstract[/h] Emerging and reemerging viruses are responsible for a number of recent epidemic outbreaks. A crucial step in predicting and controlling outbreaks is the timely and accurate characterization of emerging virus strains. We present a portable microfluidic platform containing carbon nanotube arrays with differential filtration porosity for the rapid enrichment and optical identification of viruses. Different emerging strains (or unknown viruses) can be enriched and identified in real time through a multivirus capture component in conjunction with surface-enhanced Raman spectroscopy. More importantly, after viral capture and detection on a chip, viruses remain viable and get purified in a microdevice that permits subsequent in-depth characterizations by various conventional methods. We validated this platform using different subtypes of avian influenza A viruses and human samples with respiratory infections. This technology successfully enriched rhinovirus, influenza virus, and parainfluenza viruses, and maintained the stoichiometric viral proportions when the samples contained more than one type of virus, thus emulating coinfection. Viral capture and detection took only a few minutes with a 70-fold enrichment enhancement; detection could be achieved with as little as 10[SUP]2[/SUP] EID[SUB]50[/SUB]/mL (50% egg infective dose per microliter), with a virus specificity of 90%. After enrichment using the device, we demonstrated by sequencing that the abundance of viral-specific reads significantly increased from 4.1 to 31.8% for parainfluenza and from 0.08 to 0.44% for influenza virus. This enrichment method coupled to Raman virus identification constitutes an innovative system that could be used to quickly track and monitor viral outbreaks in real time.
Copyright ? 2019 the Author(s). Published by PNAS.
[h=4]KEYWORDS:[/h] carbon nanotube; infectious disease; microfabrication; sequencing; virus isolation
PMID: 31882450 DOI: 10.1073/pnas.1910113117
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Yeh YT[SUP]1[/SUP], Gulino K[SUP]2[/SUP], Zhang Y[SUP]3[/SUP], Sabestien A[SUP]4[/SUP], Chou TW[SUP]2[/SUP], Zhou B[SUP]2[/SUP], Lin Z[SUP]3[/SUP], Albert I[SUP]4[/SUP], Lu H[SUP]5[/SUP], Swaminathan V[SUP]3[/SUP], Ghedin E[SUP]2[/SUP], Terrones M[SUP]1[/SUP].
[h=3]Author information[/h] 1 Department of Physics, The Pennsylvania State University, University Park, PA 16802; yxy155@psu.edu mut11@psu.edu. 2 Department of Biology, New York University, New York, NY 10003. 3 Department of Physics, The Pennsylvania State University, University Park, PA 16802. 4 Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA 16802. 5 Department of Veterinary and Biomedical Sciences, The Pennsylvania State University, University Park, PA 16802.
[h=3]Abstract[/h] Emerging and reemerging viruses are responsible for a number of recent epidemic outbreaks. A crucial step in predicting and controlling outbreaks is the timely and accurate characterization of emerging virus strains. We present a portable microfluidic platform containing carbon nanotube arrays with differential filtration porosity for the rapid enrichment and optical identification of viruses. Different emerging strains (or unknown viruses) can be enriched and identified in real time through a multivirus capture component in conjunction with surface-enhanced Raman spectroscopy. More importantly, after viral capture and detection on a chip, viruses remain viable and get purified in a microdevice that permits subsequent in-depth characterizations by various conventional methods. We validated this platform using different subtypes of avian influenza A viruses and human samples with respiratory infections. This technology successfully enriched rhinovirus, influenza virus, and parainfluenza viruses, and maintained the stoichiometric viral proportions when the samples contained more than one type of virus, thus emulating coinfection. Viral capture and detection took only a few minutes with a 70-fold enrichment enhancement; detection could be achieved with as little as 10[SUP]2[/SUP] EID[SUB]50[/SUB]/mL (50% egg infective dose per microliter), with a virus specificity of 90%. After enrichment using the device, we demonstrated by sequencing that the abundance of viral-specific reads significantly increased from 4.1 to 31.8% for parainfluenza and from 0.08 to 0.44% for influenza virus. This enrichment method coupled to Raman virus identification constitutes an innovative system that could be used to quickly track and monitor viral outbreaks in real time.
Copyright ? 2019 the Author(s). Published by PNAS.
[h=4]KEYWORDS:[/h] carbon nanotube; infectious disease; microfabrication; sequencing; virus isolation
PMID: 31882450 DOI: 10.1073/pnas.1910113117
Free full text