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Quantification and kinetics of viral RNA transcripts produced in Orthohantavirus infected cells
Umeå University, Faculty of Medicine, Department of Clinical Microbiology, Infectious Diseases.
Umeå University, Faculty of Science and Technology, European CBRNE Center.
Umeå University, Faculty of Science and Technology, European CBRNE Center. Umeå University, Faculty of Medicine, Department of Clinical Microbiology, Infectious Diseases.
Umeå University, Faculty of Medicine, Department of Clinical Microbiology, Virology.
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2018 (English)In: Virology Journal, ISSN 1743-422X, E-ISSN 1743-422X, Vol. 15, p. 1-7, article id 18Article in journal (Refereed) Published
Abstract [en]

Background: Rodent borne viruses of the Orthohantavirus genus cause hemorrhagic fever with renal syndrome among people in Eurasia, and hantavirus cardiopulmonary syndrome in the Americas. At present, there are no specific treatments or efficient vaccines against these diseases. Improved understanding of viral transcription and replication may instigate targeted treatment of Orthohantavirus infections. For this purpose, we investigated the kinetics and levels of viral RNA transcription during an ongoing infection in-vitro. Methods: Vero E6 cells were infected with Puumala Orthohantavirus (strain Kazan) before cells and supernatants were collected at different time points post infection for the detection of viral RNAs. A plasmid containing primer binding sites of the three Orthohantavirus segments small (S), medium (M) and large (L) was constructed and standard curves were generated to calculate the copy numbers of the individual transcripts in the collected samples. Results: Our results indicated a rapid increase in the copy number of viral RNAs after 9 h post infection. At peak days, 2-6 days after infection, the S- and M-segment transcripts became thousand and hundred-fold more abundant than the copy number of the L-segment RNA, respectively. The presence of viral RNA in the cell culture media was detected at later time-points. Conclusions: We have developed a method to follow RNA transcription in-vitro after synchronous infection of Vero cells. The obtained results may contribute to the understanding of the viral replication, and may have implications in the development of antiviral drugs targeting transcription or replication of negative stranded RNA viruses.

Place, publisher, year, edition, pages
BIOMED CENTRAL LTD , 2018. Vol. 15, p. 1-7, article id 18
Keywords [en]
Orthohantavirus, RNA segments, In-vitro infection, Quantitative real-time PCR
National Category
Microbiology in the medical area
Identifiers
URN: urn:nbn:se:umu:diva-144345DOI: 10.1186/s12985-018-0932-8ISI: 000422978500001OAI: oai:DiVA.org:umu-144345DiVA, id: diva2:1181048
Available from: 2018-02-07 Created: 2018-02-07 Last updated: 2018-06-09Bibliographically approved

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Näslund, JonasTrulsson, FredrikEvander, MagnusWesula Lwande, OliviaAhlm, ClasBucht, Göran
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