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In situ rolling circle amplification detection of Crimean Congo hemorrhagic fever virus (CCHFV) complementary and viral RNA
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology.
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2012 (English)In: Virology, ISSN 0042-6822, E-ISSN 1096-0341, Vol. 426, no 2, 87-92 p.Article in journal (Refereed) Published
Abstract [en]

Crimean Congo hemorrhagic fever virus (CCHFV) is a human pathogen that causes a severe disease with high fatality rate for which there is currently no specific treatment. Knowledge regarding its replication cycle is also highly limited. In this study we developed an in situ technique for studying the different stages during the replication of CCHFV. By integrating reverse transcription, padlock probes, and rolling circle amplification, we were able to detect and differentiate between viral RNA (vRNA) and complementary RNA (cRNA) molecules, and to detect viral protein within the same cell. These data demonstrate that CCHFV nucleocapsid protein (NP) is detectable already at 6 hours post infection in vRNA- and cRNA-positive cells. Confocal microscopy showed that cRNA is enriched and co-localized to a large extent with NP in the perinuclear area, while vRNA has a more random distribution in the cytoplasm with only some co-localize with NP. However, vRNA and cRNA did not appear to co-localize directly. 

Place, publisher, year, edition, pages
2012. Vol. 426, no 2, 87-92 p.
Keyword [en]
Crimean Congo hemorrhagic fever virus, In situ isothermal amplification, vRNA, Padlock, Colocalization
National Category
Immunology in the medical area
URN: urn:nbn:se:uu:diva-259691DOI: 10.1016/j.virol.2012.01.032ISI: 000301562900001PubMedID: 22341783OAI: oai:DiVA.org:uu-259691DiVA: diva2:845105
Available from: 2015-08-10 Created: 2015-08-10 Last updated: 2015-08-10Bibliographically approved

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