tetano
Editor, Senior Moderator
Biosens Bioelectron. 2016 Jun 15;86:150-155. doi: 10.1016/j.bios.2016.06.044. [Epub ahead of print]
[h=1]Sensitive detection of influenza viruses with Europium nanoparticles on an epoxy silica sol-gel functionalized polycarbonate-polydimethylsiloxane hybrid microchip.[/h] Liu J[SUP]1[/SUP], Zhao J[SUP]2[/SUP], Petrochenko P[SUP]3[/SUP], Zheng J[SUP]3[/SUP], Hewlett I[SUP]4[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] In an effort to develop new tools for diagnosing influenza in resource-limited settings, we fabricated a polycarbonate (PC)-polydimethylsiloxane (PDMS) hybrid microchip using a simple epoxy silica sol-gel coating/bonding method and employed it in sensitive detection of influenza virus with Europium nanoparticles (EuNPs). The incorporation of sol-gel material in device fabrication provided functionalized channel surfaces ready for covalent immobilization of primary antibodies and a strong bonding between PDMS substrates and PC supports without increasing background fluorescence. In microchip EuNP immunoassay (?ENIA) of inactivated influenza viruses, replacing native PDMS microchips with hybrid microchips allowed the achievement of a 6-fold increase in signal-to-background ratio, a 12-fold and a 6-fold decreases in limit-of-detection (LOD) in influenza A and B tests respectively. Using influenza A samples with known titers, the LOD of influenza ?ENIA on hybrid microchips was determined to be ~10[SUP]4[/SUP] TCID[SUB]50[/SUB] titer/mL and 10[SUP]3[/SUP]-10[SUP]4[/SUP] EID[SUB]50[/SUB] titer/mL. A comparison test indicated that the sensitivity of influenza ?ENIA enhanced using the hybrid microchips even surpassed that of a commercial laboratory influenza ELISA test. In addition to the sensitivity improvement, assay variation was clearly reduced when hybrid microchips instead of native PDMS microchips were used in the ?ENIA tests. Finally, infectious reference viruses and nasopharyngeal swab patient specimens were successfully tested using μENIA on hybrid microchip platforms, demonstrating the potential of this unique microchip nanoparticle assay in clinical diagnosis of influenza. Meanwhile, the tests showed the necessity of using nucleic acid confirmatory tests to clarify ambiguous test results obtained from prototype or developed point-of-care testing devices for influenza diagnosis.
Published by Elsevier B.V.
[h=4]KEYWORDS:[/h] Europium nanoparticle; Immunoassay; Influenza; Lab-on-a-chip; Sol-gel coating; Surface functionalization
PMID: 27362253 DOI: 10.1016/j.bios.2016.06.044
[PubMed - as supplied by publisher]
[h=1]Sensitive detection of influenza viruses with Europium nanoparticles on an epoxy silica sol-gel functionalized polycarbonate-polydimethylsiloxane hybrid microchip.[/h] Liu J[SUP]1[/SUP], Zhao J[SUP]2[/SUP], Petrochenko P[SUP]3[/SUP], Zheng J[SUP]3[/SUP], Hewlett I[SUP]4[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] In an effort to develop new tools for diagnosing influenza in resource-limited settings, we fabricated a polycarbonate (PC)-polydimethylsiloxane (PDMS) hybrid microchip using a simple epoxy silica sol-gel coating/bonding method and employed it in sensitive detection of influenza virus with Europium nanoparticles (EuNPs). The incorporation of sol-gel material in device fabrication provided functionalized channel surfaces ready for covalent immobilization of primary antibodies and a strong bonding between PDMS substrates and PC supports without increasing background fluorescence. In microchip EuNP immunoassay (?ENIA) of inactivated influenza viruses, replacing native PDMS microchips with hybrid microchips allowed the achievement of a 6-fold increase in signal-to-background ratio, a 12-fold and a 6-fold decreases in limit-of-detection (LOD) in influenza A and B tests respectively. Using influenza A samples with known titers, the LOD of influenza ?ENIA on hybrid microchips was determined to be ~10[SUP]4[/SUP] TCID[SUB]50[/SUB] titer/mL and 10[SUP]3[/SUP]-10[SUP]4[/SUP] EID[SUB]50[/SUB] titer/mL. A comparison test indicated that the sensitivity of influenza ?ENIA enhanced using the hybrid microchips even surpassed that of a commercial laboratory influenza ELISA test. In addition to the sensitivity improvement, assay variation was clearly reduced when hybrid microchips instead of native PDMS microchips were used in the ?ENIA tests. Finally, infectious reference viruses and nasopharyngeal swab patient specimens were successfully tested using μENIA on hybrid microchip platforms, demonstrating the potential of this unique microchip nanoparticle assay in clinical diagnosis of influenza. Meanwhile, the tests showed the necessity of using nucleic acid confirmatory tests to clarify ambiguous test results obtained from prototype or developed point-of-care testing devices for influenza diagnosis.
Published by Elsevier B.V.
[h=4]KEYWORDS:[/h] Europium nanoparticle; Immunoassay; Influenza; Lab-on-a-chip; Sol-gel coating; Surface functionalization
PMID: 27362253 DOI: 10.1016/j.bios.2016.06.044
[PubMed - as supplied by publisher]