Short communication: Stability and integrity of classical swine fever virus RNA stored at room temperature
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 ro...
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doaj-0dc4912609e44156b7ec9b5711fa69b82020-11-24T23:54:51ZengInstituto Nacional de Investigación y Tecnología Agraria y AlimentariaSpanish Journal of Agricultural Research2171-92922017-12-01153e05e0510.5424/sjar/2017153-107762567Short communication: Stability and integrity of classical swine fever virus RNA stored at room temperatureDamarys Relova0Lester J. Pérez1Liliam Ríos2Liani Coronado3Yoandry Hinojosa4Ana M. Acevedo5María T. Frías6Carmen L. Perera7National Center for Animal and Plant Health (CENSA), OIE Collaborating Centre for Diagnosis and Risk Analysis of the Caribbean Region. Ctra. de Jamaica, San José de las Lajas, La Habana 32700National Center for Animal and Plant Health (CENSA), OIE Collaborating Centre for Diagnosis and Risk Analysis of the Caribbean Region. Ctra. de Jamaica, San José de las Lajas, La Habana 32700National Center for Animal and Plant Health (CENSA), OIE Collaborating Centre for Diagnosis and Risk Analysis of the Caribbean Region. Ctra. de Jamaica, San José de las Lajas, La Habana 32700National Center for Animal and Plant Health (CENSA), OIE Collaborating Centre for Diagnosis and Risk Analysis of the Caribbean Region. Ctra. de Jamaica, San José de las Lajas, La Habana 32700National Center for Animal and Plant Health (CENSA), OIE Collaborating Centre for Diagnosis and Risk Analysis of the Caribbean Region. Ctra. de Jamaica, San José de las Lajas, La Habana 32700National Center for Animal and Plant Health (CENSA), OIE Collaborating Centre for Diagnosis and Risk Analysis of the Caribbean Region. Ctra. de Jamaica, San José de las Lajas, La Habana 32700National Center for Animal and Plant Health (CENSA), OIE Collaborating Centre for Diagnosis and Risk Analysis of the Caribbean Region. Ctra. de Jamaica, San José de las Lajas, La Habana 32700National Center for Animal and Plant Health (CENSA), OIE Collaborating Centre for Diagnosis and Risk Analysis of the Caribbean Region. Ctra. de Jamaica, San José de las Lajas, La Habana 32700Worldwide 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 37oC 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.http://revistas.inia.es/index.php/sjar/article/view/10776dried RNAdegradationvirus rescueviral diagnosisviral characterizationRNA shipment |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Damarys Relova Lester J. Pérez Liliam Ríos Liani Coronado Yoandry Hinojosa Ana M. Acevedo María T. Frías Carmen L. Perera |
spellingShingle |
Damarys Relova Lester J. Pérez Liliam Ríos Liani Coronado Yoandry Hinojosa Ana M. Acevedo María T. Frías Carmen L. Perera Short communication: Stability and integrity of classical swine fever virus RNA stored at room temperature Spanish Journal of Agricultural Research dried RNA degradation virus rescue viral diagnosis viral characterization RNA shipment |
author_facet |
Damarys Relova Lester J. Pérez Liliam Ríos Liani Coronado Yoandry Hinojosa Ana M. Acevedo María T. Frías Carmen L. Perera |
author_sort |
Damarys Relova |
title |
Short communication: Stability and integrity of classical swine fever virus RNA stored at room temperature |
title_short |
Short communication: Stability and integrity of classical swine fever virus RNA stored at room temperature |
title_full |
Short communication: Stability and integrity of classical swine fever virus RNA stored at room temperature |
title_fullStr |
Short communication: Stability and integrity of classical swine fever virus RNA stored at room temperature |
title_full_unstemmed |
Short communication: Stability and integrity of classical swine fever virus RNA stored at room temperature |
title_sort |
short communication: stability and integrity of classical swine fever virus rna stored at room temperature |
publisher |
Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria |
series |
Spanish Journal of Agricultural Research |
issn |
2171-9292 |
publishDate |
2017-12-01 |
description |
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 37oC 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. |
topic |
dried RNA degradation virus rescue viral diagnosis viral characterization RNA shipment |
url |
http://revistas.inia.es/index.php/sjar/article/view/10776 |
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