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Abstract: During the corona virus (COVID-19) pandemic, there was a sharp increase in the need for diagnostic tests that could detect the presence of SARS-CoV-2 virus or its antibodies quickly and reliably. An important type in the group of diagnostic tests are rapid antigen lateral flow immuno-assay (LFIA) tests, which operate on the immuno-chromatographic principle with the lateral flow of analyte. Clinical practice in the last year has shown that such diagnostic tests can be effective in preventing the spread of the SARS-CoV-2 virus.The development, and, thus, the production of the rapid antigen LFIA tests, is influenced by a number of factors that determine their sensitivity and accuracy indirectly. These factors are directly dependent on the type of antibody produced, which is formed as an immune response when infected with the virus. The production of the rapid antigen LFIA tests is associated with the appropriate selection of basic components that determine the type and quality of these tests. The basic components include: substrates and membranes, antigens, antibody labels and compatible buffers. The correct choice of membranes and their materials is crucial to compiling an effective rapid antigen LFIA test. This study therefore presents a comparative analysis of four commercially available SARS-CoV-2-rapid LFIA tests using state-of-the-art characterisation techniques scanning electron microscopy (SEM), inductively coupled plasma-optical emission spectrometry (ICP-OES), environmental scanning electron microscope / energy-dispersive X-ray spectroscopy (ESEM/EDX), Fourier-transform infrared spectroscopy / attenuated total reflection (FTIR/ATR) for the individual components. The obtained results were the starting point for the development and assembling of our own rapid antigen LFIA test based on gold nanoparticles as antibody labels.
Keywords: hitri antigenski testi, komponente, karakterizacija, analize, rapid antigen test, components, characterisation, analysis
Published in DKUM: 26.03.2025; Views: 0; Downloads: 3
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Abstract: The Ni/Y2O3 catalyst showed high catalytic activity. Based on this, the aim of this study was to create Ni/Y2O3 nanocomposites powder with two innovative technologies, Ultrasonic Spray Pyrolysis (USP) and lyophilisation. In the USP process, thermal decomposition of the generated aerosols in an N2/H2 reduction atmosphere caused a complete decomposition of the nickel (II) nitrate to elemental Ni, which became trapped on the formed Y2O3 nanoparticles. The Ni/Y2O3 nanocomposite particles were captured via gas washing in an aqueous solution of polyvinylpyrrolidone (PVP) in collection bottles. PVP was chosen for its ability to stabilise nano-suspensions and as an effective cryoprotectant. Consequently, there was no loss or agglomeration of Ni/Y2O3 nanocomposite material during the lyophilisation process. The Ni/Y2O3 nanocomposite powder was analysed using ICP-MS, SEM-EDX, and XPS, which showed the impact of different precursor concentrations on the final Ni/Y2O3 nanocomposite particle composition. In a final step, highly concentrated Ni/Y2O3 nanocomposite ink (Ni/Y2O3 > 0.140 g/mL) and test coatings from this ink were prepared by applying them on a white matte photo paper sheet. The reflection curve of the prepared Ni/Y2O3 nanocomposite coating showed a local maximum at 440 nm with a value of 39% reflection. Given that Ni is located on the surface of the Ni/Y2O3 nanocomposite in the elemental state and according to the identified properties, tests of the catalytic properties of this coating will be performed in the future.
Keywords: Ultrasonic Spray Pyrolysis (USP), lyophilisation, Ni/Y2O3 nanocomposite, characterisation, coatings
Published in DKUM: 20.03.2025; Views: 0; Downloads: 5
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Abstract: The manuscript presents the optical properties of directly deposited films of gold nanoparticles (AuNPs) prepared by the Ultrasonic Spray Pyrolysis (USP) technology. Four samples were produced, with AuNP deposition times on the glass substrate of 15 min, 30 min, 1 h and 4 h. The morphological characterisation of the deposited films showed that the size of the first deposited AuNPs was between 10 and 30 nm, while, with a longer duration of the deposition process, larger clusters of AuNPs grew by coalescence and aggregation. The prepared layers were characterised optically with Ultraviolet–visible spectroscopy (UV–vis) and ellipsometry. The ellipsometric measurements showed an increasingly denser and thicker effective thickness of the AuNP layers. The extinction spectra displayed a clear local surface plasmonic resonance (LSPR) signature (peak 520–540 nm), indicating the presence of isolated particles in all the samples. For all AuNP layers, the imaginary part of the parallel and perpendicular components of the anisotropic dielectric function was dominated by a central peak at around 2.2 eV, corresponding to the LSPR of isolated particles, and a high-energy shoulder due to Au interband transitions. It was shown that, as the density of particles increased, the extinction cross-section grew over the whole spectral range where measurements are taken. Thus, the response can be explained with an enhanced electromagnetic response between the AuNPs that can be connected to the increase in particle density, but also by the formation of clusters and irregular structures.
Keywords: optical characteristics, gold nanoparticles’ films, UV–vis, ellipsometry measurements
Published in DKUM: 28.05.2024; Views: 124; Downloads: 0

Abstract: V doktorskem delu so predstavljene študije, eksperimentalno delo, tehnike karakterizacije, rezultati in analize, s pomočjo katerih smo potrdili hipotezo, da je možno Au nanodelce (AuND) pripravljene z metodo ultrazvočne razpršilne pirolize (USP), konjugirati s ciljnimi antigeni SARS-CoV-2 in rekombinantnim IgG, da bodo delovali v imunoserološkem hitrem testu na lateralni tok (LFIA), za potrditev potencialne prisotnosti protiteles proti virusu SARS-CoV-2. S ciljno izbranimi parametri (0,5 g/l Au, 5 l/min N2, 2,5 l/min H2) smo z USP sintetizirali AuND iz prekurzorja na osnovi zlatovega (III) klorida. AuND smo zbirali v steklenicah z demineralizirano vodo. Določili smo optimalne stabilizatorje in njihove koncentracije: 2,5 g/L PVP 40K ((C6H9NO)n) ter naknadno dodani 1 g/L trinatrijevega citrata (Na3C6H5O7) in saharoze (C12H22O11), da bi preprečili AuND aglomeracijo. Presevna elektronska mikroskopija AuND je pokazala, da so okrogli, s povprečno velikostjo okoli 50 nm in ozko velikostno porazdelitvijo, brez defektov in nizkim ZETA potencialom, 2,4 mV, pri pH 7,5. Za odstranitev vode iz suspenzije z AuND in stabilizatorji je bila uporabljena liofiliazcija, ki je bila izvedena pri naslednjih pogojih: zamrzovanje pri –40 °C, 4 h, atmosferski tlak; primarno sušenje pri +20 °C, 12 h, 1–4 Pa; sekundarno sušenje pri +30 °C, 33 h, 1–4 Pa. Nastali posušeni AuND so ohranili vse lastnosti, ki so potrebne za funkcijo oznak v hitrih LFIA testih: visoka površinska energija, površinska plazmonska resonanca in proste karboksilne skupine za konjugacijo. V nadaljevanju izvajanja eksperimentov smo, da bi potrdili doktorske hipoteze, izvedli raziskavo in optimizacijo konjugacije AuND z antigenom SARS-CoV-2, ki je bodičasti protein S1 (ak 1-681), in s protitelesi na zajčji IgG (RMG03). Za določitev poteka konjugacije AuND smo uporabili posredne (gelske elektroforeze, UV-Vis, preizkus delovanja) in direktne (XPS, SIMS) metode. Rezultati so pokazali, da se med našim postopkom konjugacije, v fazah čiščenja odstranimo večinski delež PVP in saharoze, ter s tem sprostimo proste karboksilne funkcionalne skupine citratnih ionov, ki so vezani na povšini AuND. Na teh prostih mestih lahko poteče konjugacija prek prostih -NH2 skupin na proteinih. Njihovo funkcionalnost smo ovrednotili z eksperimentalnimi testiranji lastno razvitega LFIA testa. Na osnovi tega smo postavili model in mehanizem konjugacije AuND, ki predpostavlja, da poteka konjugacija z antigeni oziroma protitelesi prek tvorbe peptidnih vezi med N-koncem proteina in prostimi karboksilnimi skupinami citratnega iona, ki je vezan na površino AuND prek svojih karboksilnih skupin. Za dodatno potrditev funkcionalizacije AuND z izbranimi biomolekulami smo v sklepnem delu doktorskega dela izvedli klinično študijo na pacientih UKC MB, ki je bila odobrena s sklepom KME RS št. 0120-148/2021/3. Rezultati študije so pokazali, da testi dosegajo visoko občutljivost (83 %) tako na vzorcih nosnega sluza kot na vzorcih seruma, slabšo specifičnost (66 % sluz, 74 % serum), visoko negativno napovedno vrednost (94 %) in slabšo pozitivno napovedno vrednost (38 % sluz, 45 % serum). S študijo smo potrdili delovanje AuND v funkciji oznak, saj so se testne in kontrolne linije v lastno razvitem LFIA hitrem testu obarvale.
Keywords: Nanodelci zlata, SARS-CoV-2 virus, stabilizacija, konjugacija, LFIA testi, karakterizacija
Published in DKUM: 28.03.2024; Views: 347; Downloads: 58
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Keywords: rapid antigen tests, recycling, characterization, nanogold, plastic
Published in DKUM: 12.02.2024; Views: 383; Downloads: 6