Here, we describe a highly sensitive method that allows for the correct quantification of inhibition effect with a higher degree of accuracy directly at the molecular level. The protocol involves two stages, namely serological virus titration in comparison with the same procedure for virus-effector mixture. Owing to the robustness of the analysis this assay can be performed on crude cellular and plant extracts, and therefore it may be suitable for the routine analysis of clinical samples, or in the field. The efficiency of the approach to the quantification of the inhibition effect of polysaccharide glucuronoxylomannan (GXM) on the infection efficiency of the tobacco mosaic virus (TMV) was investigated using advanced serological approaches based on label-free surface plasmon resonance technique. It was shown that GXM drastically decreases the efficiency of TMV infection by blocking up to 70% of the virus shell. The obtained results are in conformity with the method of indicator plant infection.
The nanosized composites based on the natural polysaccharides and nanoparticles of noble metals are promising candidates for efficient antiviral drugs. However, the complexity of such objects, their diversity and novelty necessitate the development of new analytical methods for investigation of such supramolecular architectures. In this work, which was recently developed for SPR-based instrumentation, the concept of variative refraction (DViFA, density variations in fixed architectures) was used to elucidate the mechanism of the antiviral action of a polysaccharide with gold nanoparticles grown in it. The SPR data were confirmed by direct biological tests: the effect of the native polysaccharide glucuronoxylomannan (GXM) obtained from the fungus Ganoderma adspersum and gold nanocomposites thereon on the infection of Datura stramonium with tobacco mosaic virus (TMV) was investigated. Both drugs suppress the development of viral infections. However, if for high concentrations the characteristic activity of the composite is somewhat lower than for GXM, then with an increase in dilution, the effectiveness of the composite increases significantly, up to a twofold excess. It has been reasonably suggested that the mechanism of antiviral action is associated with the formation of clusters of viruses that are no longer capable of infecting cells.
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