Quantitative in situ hybridization using strand specific RNA probes: expression of the bunyavirus Germiston S segment in mosquito cells.

Abstract

Infection of Vero (monkey) cells by Germiston bunyavirus is highly cytopathic with cell lysis and virus production at a high titre, whereas infection of Aedes albopictus C6/36 (mosquito) cells leads, after an acute primary phase, to a persistent non-cytopathic infection with a loss in virus production. In this report we demonstrate that single-stranded RNA probes can be successfully used in an in situ hybridization assay to quantify viral expression during this persistent infection. The steady-state levels of viral S-RNA segment (genomic and messenger sense) during the acute phase were similar to those observed in lytically infected Vero cells, but appeared delayed. Both senses of S-RNA were detected throughout persistent infection but in lower amounts, in less than 10% of the cells and always in the cytoplasm of infected cells. The number of copies per cell of messenger sense S-RNAs remained low during persistent infection whereas a higher fluctuation was observed for genomic S-RNAs. In situ hybridization with specific stranded RNA probes provides both qualitative and quantitative informations, that can lead to a better understanding of virus-cell interactions.

• Authors: B Delord, J D Poveda, T Astier-Gin, S Gerbaud, J P Wattiaux, H J Fleury
• Journal: Molecular and cellular probes (Mol Cell Probes) Vol. 4 Issue 4 Pg. 247-59 (Aug 1990) ISSN: 0890-8508 [Print] England
• PMID: 2402248 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• RNA Probes
• RNA, Viral

Topics

• Aedes
• Animals
• Bunyaviridae (genetics, growth & development, physiology)
• Cell Line
• Nucleic Acid Hybridization
• RNA Probes
• RNA, Viral (analysis, genetics)
• Vero Cells
• Virus Replication (genetics)