BackgroundThe timely and accurate diagnosis of specific influenza virus strains is crucial to effective prophylaxis, vaccine preparation and early antiviral therapy. The detection of influenza A viruses is mainly accomplished using polymerase chain reaction (PCR) techniques or antibody-based assays. In conjugation with the immunoassay utilizing monoclonal antibody, mass spectrometry is an alternative to identify proteins derived from a target influenza virus. Taking advantage of the large surface area-to-volume ratio, antibody-conjugated magnetic nanoparticles can act as an effective probe to extract influenza virus for sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) and on-bead mass spectrometric analysis.ResultsIron oxide magnetic nanoparticles (MNP) were functionalized with H5N2 viral antibodies targeting the hemagglutinin protein and capped with methoxy-terminated ethylene glycol to suppress nonspecific binding. The antibody-conjugated MNPs possessed a high specificity to H5N2 virus without cross-reactivity with recombinant H5N1 viruses. The unambiguous identification of the captured hemagglutinin on magnetic nanoparticles was realized by SDS-PAGE visualization and peptide sequence identification using liquid chromatography-tandem mass spectrometry (LC-MS/MS).ConclusionsThe assay combining efficient magnetic separation and MALDI-MS readout offers a rapid and sensitive method for virus screening. Direct on-MNP detection by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) provided high sensitivity (~103 EID50 per mL) and a timely diagnosis within one hour. The magnetic nanoparticles encapsulated with monoclonal antibodies could be used as a specific probe to distinguish different subtypes of influenza.
We have applied a reusable silicon nanowire field-effect transistor (SiNW-FET) as a biosensor to conduct ultrasensitive detection of H5N2 avian influenza virus (AIV) in very dilute solution. The reversible surface functionalization of SiNW-FET was made possible using a disulfide linker. In the surface functionalization, 3-mercaptopropyltrimethoxysilane (MPTMS) was first modified on the SiNW-FET (referred to as MPTMS/SiNW-FET), with subsequent dithiothreitol washing to reduce any possible disulfide bonding between the thiol groups of MPTMS. Subsequently, receptor molecules could be immobilized on the MPTMS/SiNW-FET by the formation of a disulfide bond. The success of the reversible surface functionalization was verified with fluorescence examination and electrical measurements. A surface topograph of the SiNW-FET biosensor modified with a monoclonal antibody against H5N2 virus (referred to as mAb(H5)/SiNW-FET) after detecting approximately 10(-17) M H5N2 AIVs was scanned by atomic force microscopy to demonstrate that the SiNW-FET is capable of detecting very few H5N2 AIV particles.
In this report, an amorphous indium gallium zinc oxide (a-IGZO) thin-film was fabricated by DC-sputtering at different oxygen partial pressure values. The transmittance of the thin films could be enhanced by treated with oxygen during sputtering and greater than 90% at 300~700 nm. It has been found that the oxide semiconductor properties are highly dependent on oxygen content, because oxygen vacancies provide the needed free carriers for electrical conduction. Therefore, the conductivity could be controlled by adjusting the flux of the mixture oxygen during film deposition. With the increasing amount of oxygen, the IGZO thin film would change from conductor to semiconductor, and finally become an insulator. During this tunable characteristic, one can adjust the resistivity according to different kinds of IGZOs, such as InGaZnO 4 we can get an amorphous, transparent thin film with the mobility around 6 cm 2 /V-s and the resistivity around 106 ohm when the oxygen to argon ratio is 0.66%.
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