title: In situ rolling circle amplification detection of Crimean Congo hemorrhagic fever virus (CCHFV) complementary and viral RNA.
abstract: 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.
authors.affiliation: Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, SE-171 77 Stockholm, Sweden.
authors.forename: Cecilia
authors.initials: C
authors.lastname: Andersson
authors.forename: Sara
authors.initials: S
authors.lastname: Henriksson
authors.forename: Karl-Eric
authors.initials: KE
authors.lastname: Magnusson
authors.forename: Mats
authors.initials: M
authors.lastname: Nilsson
authors.forename: Ali
authors.initials: A
authors.lastname: Mirazimi
mesh_terms: D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D003225:Hemorrhagic Fever Virus, Crimean-Congo / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology; D006479:Hemorrhagic Fever, Crimean / Q000175:diagnosis / Q000821:virology*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D018075:RNA, Complementary / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics*; D014709:Vero Cells; D014779:Virus Replication
chemical_list: D018075:RNA, Complementary; D012367:RNA, Viral