Using a Biosensor Based on Surface-Enhanced Raman Scattering to Identify Influenza Viruses in Biological Fluids

Abstract

Biosensors based on the effect of surface-enhanced Raman scattering obtained on silver nanoclusters modified with DNA aptamers allow viruses to be detected with high sensitivity. However, measurements in biological media are complicated by the nonspecific sorption of biomolecules on silver. Conditions for preparing samples of biological fluids that allow the nonspecific sorption of biomolecules to be nullified are studied.

A Combination of Membrane Filtration and Raman-Active DNA Ligand Greatly Enhances Sensitivity of SERS-Based Aptasensors for Influenza A Virus

“…Recently, we have demonstrated the efficiency of several sensors based on the RHA0385 aptamer ( Kukushkin et al, 2019 ; Gribanev et al, 2021 ; Gribanyov et al, 2021 ; Zhdanov et al, 2022 ). Here, we provided data about the selectivity of the aptamer toward influenza A virus and assembling of sandwich-like structures to verify the hypothetic model of the sensor.…”

“…Sandwich assay was performed similar to our previous work ( Zhdanov et al, 2022 ). In particular, functionalization of silver nanoparticles with the aptamer was performed in the following way.…”

“…In particular, functionalization of silver nanoparticles with the aptamer was performed in the following way. A measure of 300 µl of viruses (influenza A virus, influenza B virus, Newcastle disease virus or allantoic fluid) diluted in phosphate-buffered saline was incubated with 67 nM of the BODIPY FL-labeled aptamer (concentration of this oligonucleotide was two times higher than the previous setup) ( Zhdanov et al, 2022 ). The solution was mixed with 200 µl of functionalized silver nanoparticles and incubated for 1 min (the time was decreased 4 times compared to our previous work) ( Zhdanov et al, 2022 ).…”

“…A measure of 300 µl of viruses (influenza A virus, influenza B virus, Newcastle disease virus or allantoic fluid) diluted in phosphate-buffered saline was incubated with 67 nM of the BODIPY FL-labeled aptamer (concentration of this oligonucleotide was two times higher than the previous setup) ( Zhdanov et al, 2022 ). The solution was mixed with 200 µl of functionalized silver nanoparticles and incubated for 1 min (the time was decreased 4 times compared to our previous work) ( Zhdanov et al, 2022 ). SERS was acquired using a RaPort instrument (Enhanced Spectrometry, United States) with a laser radiation wavelength of 532 nm, and an operating power of 30 mW was used for registration.…”

“…Our previous efforts produced a SERS-based aptasensor on the solid substrate with an LoD of 5 10 –4 HAU/ml that corresponds to 8 10 4 VP/ml ( Kukushkin et al, 2019 ) and a SERS-based aptasensor on the aggregated colloid silver nanoparticles with an LoD of 2 10 5 VP/ml ( Gribanev et al, 2021 ; Gribanyov et al, 2021 ; Zavyalova et al, 2021 ; Zhdanov et al, 2022 ); both were based on the RHA0385 aptamer to hemagglutinin of the influenza A virus ( Shiratori et al, 2014 ). Despite being less sensitive and inferior in a range of virus determination, the latter is preferred because colloid nanoparticles are much cheaper and easier in the production not requiring sophisticated equipment and silicon substrates.…”