Assuntos
COVID-19/complicações , COVID-19/imunologia , Pérnio/imunologia , Pérnio/virologia , Pneumonia Viral/complicações , Pneumonia Viral/imunologia , Adolescente , COVID-19/diagnóstico , Criança , Pré-Escolar , Feminino , França , Humanos , Lactente , Masculino , Pandemias , Pneumonia Viral/diagnóstico , Pneumonia Viral/virologia , SARS-CoV-2RESUMO
Amoeba-infecting giant viruses are recently discovered viruses that have been isolated from diverse environments all around the world. In parallel to isolation efforts, metagenomics confirmed their worldwide distribution from a broad range of environmental and host-associated samples, including humans, depicting them as a major component of eukaryotic viruses in nature and a possible resident of the human/animal virome whose role is still unclear. Nevertheless, metagenomics data about amoeba-infecting giant viruses still remain scarce, mainly because of methodological limitations. Efforts should be pursued both at the metagenomic sample preparation level and on in silico analyses to better understand their roles in the environment and in human/animal health and disease.
Assuntos
Amoeba/virologia , Vírus Gigantes/classificação , Vírus Gigantes/genética , Metagenômica/métodos , Animais , DNA Viral/genética , Genoma Viral/genética , Vírus Gigantes/isolamento & purificação , HumanosRESUMO
Here we describe an optimized molecular protocol for the universal detection and identification of flaviviruses. It combines the convenient real-time polymerase chain reaction (PCR) format with a broad spectrum of flavivirus detection. This assay, based on the amplification of a 269-272 nt (depending on the flavivirus tested) region at the N terminal end of the NS5 gene, enabled the amplification of 51 flavivirus species and 3 tentative species. Sequencing of the amplicons produced by reverse transcriptase (RT)-PCR permitted the reliable taxonomic identification of flavivirus species by comparison with reference sequences available in databases, using either the BLASTN algorithm or a simple phylogenetic reconstruction. The limit of detection of the assay (2-20,500 copies/reaction depending on the virus tested) allowed the detection of different flaviviruses from a series of human sera or veterinary samples. Altogether, the characteristics of this technique make it a good candidate for the identification of previously identified flaviviruses in cell culture and the investigation of field samples, and also a promising tool for the discovery and identification of new species, including viruses distantly related to "classical" arthropod-borne flaviviruses.
