NDLI: Quantification and kinetics of viral RNA transcripts produced in Orthohantavirus infected cells

Content Provider	Springer Nature : BioMed Central
Author	Wigren Byström, Julia Näslund, Jonas Trulsson, Fredrik Evander, Magnus Wesula Lwande, Olivia Ahlm, Clas Bucht, Göran
Abstract	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.
Related Links	https://virologyj.biomedcentral.com/counter/pdf/10.1186/s12985-018-0932-8.pdf
Ending Page	7
Page Count	7
Starting Page	1
File Format	HTM / HTML
DOI	10.1186/s12985-018-0932-8
Journal	Virology Journal
Issue Number	1
Volume Number	15
Language	English
Publisher	BioMed Central
Publisher Date	2018-01-19
Access Restriction	Open
Subject Keyword	Virology Orthohantavirus RNA segments In-vitro infection Quantitative real-time PCR
Content Type	Text
Resource Type	Article
Subject	Virology Infectious Diseases
Journal Impact Factor	4/2023
5-Year Journal Impact Factor	3.8/2023

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