Stability and integrity of classical swine fever virus RNA stored at room temperature

Damarys Relova, Lester J. Pérez, Liliam Ríos, Liani Coronado, Yoandry Hinojosa, Ana M. Acevedo, María T. Frías, Carmen L. Perera

Research output: Contribution to journalArticlepeer-review

Abstract

Worldwide cooperation between laboratories working with classical swine fever virus (CSFV) requires exchange of virus isolates. For this purpose, shipment of CSFV RNA is a safe alternative to the exchange of infectious material. New techniques using desiccation have been developed to store RNA at room temperature and are reported as effective means of preserving RNA integrity. In this study, we evaluated the stability and integrity of dried CSFV RNA stored at room temperature. First, we determined the stability of CSFV RNA covering CSFV genome regions used typically for the detection of viral RNA in diagnostic samples by reverse transcription-polymerase chain reaction (RT-PCR). To this end, different concentrations of in vitro-transcribed RNAs of the 5'-untranslated region and of the NS5B gene were stored as dried RNA at 4, 20, and 37°C for two months. Aliquots were analyzed every week by CSFV-specific quantitative real-time RT-PCR. Neither the RNA concentration nor the storage temperature did affect CSFV RNA yields at any of the time evaluated until the end of the experiment. Furthermore, it was possible to recover infectious CSFV after transfection of SK-6 cells with dried viral RNA stored at room temperature for one week. The full-length E2 of CSFV was amplified from all the recovered viruses, and nucleotide sequence analysis revealed 100% identity with the corresponding sequence obtained from RNA of the original material. These results show that CSFV RNA stored as dried RNA at room temperature is stable, maintaining its integrity for downstream analyses and applications.

Original languageEnglish (US)
Article numbere05SC03
JournalSpanish Journal of Agricultural Research
Volume15
Issue number3
DOIs
StatePublished - 2017
Externally publishedYes

Keywords

  • Degradation
  • Dried RNA
  • RNA shipment
  • Viral characterization
  • Viral diagnosis
  • Virus rescue

ASJC Scopus subject areas

  • Agronomy and Crop Science

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