Analysis of Negative-Strand RNA Viruses by RT-qPCR: Rift Valley Fever Virus and Toscana Virus

Methods Mol Biol. 2024:2824:67-80. doi: 10.1007/978-1-0716-3926-9_5.

Abstract

RT-qPCR allows the detection of viruses and the monitoring of viral replication. This technique was extensively employed during the SARS-CoV-2 pandemic, where it demonstrated its efficiency and robustness. Here we describe the analysis of Rift Valley fever and Toscana virus infections over time, achieved through the RT-qPCR quantification of the viral genome. We further elaborate on the method to discriminate between genomic and antigenomic viral RNAs by using primers specific for each strand during the reverse transcription step.

Keywords: Bunyavirus; Detection method; Negative-strand RNA virus; RT-qPCR; Rift Valley fever virus; Toscana virus.

MeSH terms

  • Animals
  • Genome, Viral
  • Humans
  • RNA, Viral* / genetics
  • Real-Time Polymerase Chain Reaction / methods
  • Rift Valley Fever* / diagnosis
  • Rift Valley Fever* / virology
  • Rift Valley fever virus* / genetics
  • Virus Replication / genetics

Substances

  • RNA, Viral