Graphene Oxide Conjugated with Trizol Reagent, Magnetic Beads and Iron Nanoparticles for RNA Extraction Methods of Rabies Virus
Rabies is a disease caused by Rabies virus (RABV - Rabies Lyssavirus) which is an RNA
virus of the genus Lyssavirus with a long history of infection and is endowed with great harm
to humanity. In addition to vaccination, surveillance is one of the main means of rabies
control whose diagnosis is mainly obtained by molecular biology techniques through
ribonucleic acid (RNA) extraction, complementary DNA synthesis and polymerase chain
reaction (RT-PCR) which depend on several reagents. Thus, the search for alternative
methods of extraction that are more efficient and cost-efficient are essential to cause a
direct impact on strengthening rabies surveillance and control. The usual protocols start
with the extraction of RNA from the samples using Trizol or by magnetic beads and then
proceed to RT-PCR. In order to increase the efficiency of extraction methods, nanoscience is
an important tool with some carbon materials as graphene oxides (GO) that are interesting
due to the oxygen levels incorporated for potential biotechnologies applications. In this
work, we report different methods using combinations of GO with Trizol, magnetic beads
and iron oxide nanoparticles to extract RNA of RABV. The BHK-21 cells infected with Pasteur
Virus (PV) were used. The GO were sintesited by the Hummers Modificated method and
characterized. The GO was used in the extraction methods and the efficiency was measured
by a RNA quantification using a spectrophotometer. The influence of the alternative
methods with GO were determined with RT-PCR and agarose gel electrophoresis. Graphene
oxide presents phase separation on the Trizol RNA extraction method but it only had a
promising interaction with the magnetic beads and with iron nanoparticles which improve
the quantity of RNA material extracted, showing these systems as a low cost and high
efficiency alternative method for RNA extraction of RABV.