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In situ rolling circle amplification detection of Crimean Congo hemorrhagic fever virus (CCHFV) complementary and viral RNA
Swedish Institute Communicable Disease Control.
Uppsala University.
Linköping University, Department of Clinical and Experimental Medicine, Medical Microbiology. Linköping University, Faculty of Health Sciences.
Uppsala University.
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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
Elsevier , 2012. Vol. 426, no 2, 87-92 p.
Keyword [en]
Crimean Congo hemorrhagic fever virus, In situ isothermal amplification, vRNA, Padlock, Colocalization
National Category
Medical and Health Sciences
URN: urn:nbn:se:liu:diva-76521DOI: 10.1016/j.virol.2012.01.032ISI: 000301562900001OAI: diva2:515148
Funding Agencies|Swedish Medical Research Council|K2010-57X-0349-01-3|Available from: 2012-04-12 Created: 2012-04-11 Last updated: 2012-04-12

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Magnusson, Karl-Eric
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