Detection of Viral −RNA and +RNA Strands in Enterovirus-Infected Cells and Tissues
| Autor: | Varpu Marjomäki, Anni Honkimaa, Mira Laajala, Heikki Hyöty, Kerttu Kalander, Sami Oikarinen, Sami Salmikangas, Iryna Shanina, Jutta E. Laiho, Marc S. Horwitz |
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| Přispěvatelé: | Tampere University, BioMediTech, Clinical Medicine |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Microbiology (medical) replication 030106 microbiology Cell In situ hybridization Biology branched DNA medicine.disease_cause infektiot Microbiology Article antiviral drugs 03 medical and health sciences chemistry.chemical_compound positive RNA Virology medicine BDNA test lcsh:QH301-705.5 replikaatio enterovirus virus diseases RNA Translation (biology) negative RNA enterovirukset 030104 developmental biology medicine.anatomical_structure lcsh:Biology (General) chemistry Virion assembly Enterovirus in situ hybridization 3111 Biomedicine DNA |
| Zdroj: | Microorganisms, Vol 8, Iss 1928, p 1928 (2020) Microorganisms Volume 8 Issue 12 |
| ISSN: | 2076-2607 |
| DOI: | 10.3390/microorganisms8121928 |
| Popis: | The current methods to study the distribution and dynamics of viral RNA molecules inside infected cells are not ideal, as electron microscopy and immunohistochemistry can only detect mature virions, and quantitative real-time PCR does not reveal localized distribution of RNAs. We demonstrated here the branched DNA in situ hybridization (bDNA ISH) technology to study both the amount and location of the emerging &minus RNA and +RNA during acute and persistent enterovirus infections. According to our results, the replication of the viral RNA started 2&ndash 3 h after infection and the translation shortly after at 3&ndash 4 h post-infection. The replication hotspots with newly emerging &minus RNA were located quite centrally in the cell, while the +RNA production and most likely virion assembly took place in the periphery of the cell. We also discovered that the pace of replication of &minus RNA and +RNA strands was almost identical, and &minus RNA was absent during antiviral treatments. ViewRNA ISH with our custom probes also showed a good signal during acute and persistent enterovirus infections in cell and mouse models. Considering these results, along with the established bDNA FISH protocol modified by us, the effects of antiviral drugs and the emergence of enterovirus RNAs in general can be studied more effectively. |
| Databáze: | OpenAIRE |
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