Abstract
Virological analysis is time-consuming and expensive. The aim of this work is to demonstrate the applicability of laser-induced fluorescence (LIF) to the classification of viruses, reducing the time for this analysis and its costs. Experimental tests were performed in which different viruses were irradiated with a UV laser emitting at 266 nm and the emitted spectra were recorded by a spectrometer. The classification techniques show the possibility of discriminating viruses. Although the application of the LIF technique to biological agents has been thoroughly studied by many researchers over the years, this work aims at validating for the first time its applicability to virological analyses. The development of a fast virological analysis may revolutionize this field, allowing fast responses to epidemiologic events, reducing their risks and improving the efficiency of monitoring environments. Moreover, a cost reduction may lead to an increase in the monitoring frequency, with an obvious enhancement of safety and prevention.
Highlights
The measurement of laser-induced fluorescence on viruses has been investigated, and the results show the interesting potential of this technique
The laser-induced fluorescence (LIF) approach is based on exciting the fluorophores contained in the envelope proteins of the viruses by proper radiation
The classification is allowed by a proper classification algorithm, such as the three shown in this work
Summary
Once the volume to be analysed has been reduced, the test phase can be carried out by virus isolation on a susceptible cell line or through innovative molecular methods based on PCR, including qRT-PCR1, RT-PCR combined with the hybridization to specific probes[2], nanofluidic real-time RT-PCR, digital PCR, etc.[3,4,5] All these approaches show several limitations related to the nature of the viruses and the environment where they have been collected. The isolation of wild-type hepatitis A virus on specific cell lines (if the particular strain can be cultured, for instance, wt HM-175 strain9) may take 3–5 weeks and at least two passages on cells, whereas the molecular methods require 3–5 days[10] For this reason, drinking water analyses provide biological (bacteria) and chemical details, while virological analysis is avoided until an outbreak spreads. By irradiating the sample with UV light, some molecules absorb and re-emit less www.nature.com/scientificreports/
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