Impact of RNA Degradation on Viral Diagnosis: An Understated but Essential Step for the Successful Establishment of a Diagnosis Network
The current global conditions, which include intensive globalization, climate changes, and viral evolution among other factors, have led to an increased emergence of viruses and new viral diseases; RNA viruses are key drivers of this evolution. Laboratory networks that are linked to central referenc...
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| Format: | Article |
| Language: | English |
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MDPI AG
2018-02-01
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| Series: | Veterinary Sciences |
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| Online Access: | http://www.mdpi.com/2306-7381/5/1/19 |
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| author | Damarys Relova Liliam Rios Ana M. Acevedo Liani Coronado Carmen L. Perera Lester J. Pérez |
| author_facet | Damarys Relova Liliam Rios Ana M. Acevedo Liani Coronado Carmen L. Perera Lester J. Pérez |
| author_sort | Damarys Relova |
| collection | DOAJ |
| description | The current global conditions, which include intensive globalization, climate changes, and viral evolution among other factors, have led to an increased emergence of viruses and new viral diseases; RNA viruses are key drivers of this evolution. Laboratory networks that are linked to central reference laboratories are required to conduct both active and passive environmental surveillance of this complicated global viral environment. These tasks require a continuous exchange of strains or field samples between different diagnostic laboratories. The shipment of these samples on dry ice represents both a biological hazard and a general health risk. Moreover, the requirement to ship on dry ice could be hampered by high costs, particularly in underdeveloped countries or regions located far from each other. To solve these issues, the shipment of RNA isolated from viral suspensions or directly from field samples could be a useful way to share viral genetic material. However, extracted RNA stored in aqueous solutions, even at −70 °C, is highly prone to degradation. The current study evaluated different RNA storage conditions for safety and feasibility for future use in molecular diagnostics. The in vitro RNA-transcripts obtained from an inactivated highly pathogenic avian influenza (HPAI) H5N1 virus was used as a model. The role of secondary structures in the protection of the RNA was also explored. Of the conditions evaluated, the dry pellet matrix was best able to protect viral RNA under extreme storage conditions. This method is safe, cost-effective and assures the integrity of RNA samples for reliable molecular diagnosis. This study aligns with the globally significant “Global One Health” paradigm, especially with respect to the diagnosis of emerging diseases that require confirmation by reference laboratories. |
| first_indexed | 2024-12-14T22:12:17Z |
| format | Article |
| id | doaj.art-d2eca25faf474b838c59027351edd0d6 |
| institution | Directory Open Access Journal |
| issn | 2306-7381 |
| language | English |
| last_indexed | 2024-12-14T22:12:17Z |
| publishDate | 2018-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Veterinary Sciences |
| spelling | doaj.art-d2eca25faf474b838c59027351edd0d62022-12-21T22:45:43ZengMDPI AGVeterinary Sciences2306-73812018-02-01511910.3390/vetsci5010019vetsci5010019Impact of RNA Degradation on Viral Diagnosis: An Understated but Essential Step for the Successful Establishment of a Diagnosis NetworkDamarys Relova0Liliam Rios1Ana M. Acevedo2Liani Coronado3Carmen L. Perera4Lester J. Pérez5Centro Nacional de Sanidad Agropecuaria (CENSA), OIE Collaborating Centre for Diagnosis and Risk Analysis of the Caribbean Region, La Habana 32700, CubaReiman Cancer Research Laboratory, Faculty of Medicine, University of New Brunswick, Saint John, NB E2L 4L5, CanadaCentro Nacional de Sanidad Agropecuaria (CENSA), OIE Collaborating Centre for Diagnosis and Risk Analysis of the Caribbean Region, La Habana 32700, CubaCentro Nacional de Sanidad Agropecuaria (CENSA), OIE Collaborating Centre for Diagnosis and Risk Analysis of the Caribbean Region, La Habana 32700, CubaCentro Nacional de Sanidad Agropecuaria (CENSA), OIE Collaborating Centre for Diagnosis and Risk Analysis of the Caribbean Region, La Habana 32700, CubaDalhousie Medicine New Brunswick, Dalhousie University, Saint John, NB E2L 4L5, CanadaThe current global conditions, which include intensive globalization, climate changes, and viral evolution among other factors, have led to an increased emergence of viruses and new viral diseases; RNA viruses are key drivers of this evolution. Laboratory networks that are linked to central reference laboratories are required to conduct both active and passive environmental surveillance of this complicated global viral environment. These tasks require a continuous exchange of strains or field samples between different diagnostic laboratories. The shipment of these samples on dry ice represents both a biological hazard and a general health risk. Moreover, the requirement to ship on dry ice could be hampered by high costs, particularly in underdeveloped countries or regions located far from each other. To solve these issues, the shipment of RNA isolated from viral suspensions or directly from field samples could be a useful way to share viral genetic material. However, extracted RNA stored in aqueous solutions, even at −70 °C, is highly prone to degradation. The current study evaluated different RNA storage conditions for safety and feasibility for future use in molecular diagnostics. The in vitro RNA-transcripts obtained from an inactivated highly pathogenic avian influenza (HPAI) H5N1 virus was used as a model. The role of secondary structures in the protection of the RNA was also explored. Of the conditions evaluated, the dry pellet matrix was best able to protect viral RNA under extreme storage conditions. This method is safe, cost-effective and assures the integrity of RNA samples for reliable molecular diagnosis. This study aligns with the globally significant “Global One Health” paradigm, especially with respect to the diagnosis of emerging diseases that require confirmation by reference laboratories.http://www.mdpi.com/2306-7381/5/1/19viral diagnosisRNA storagesecondary structureslaboratory networkssample exchangereal time RT-PCR |
| spellingShingle | Damarys Relova Liliam Rios Ana M. Acevedo Liani Coronado Carmen L. Perera Lester J. Pérez Impact of RNA Degradation on Viral Diagnosis: An Understated but Essential Step for the Successful Establishment of a Diagnosis Network Veterinary Sciences viral diagnosis RNA storage secondary structures laboratory networks sample exchange real time RT-PCR |
| title | Impact of RNA Degradation on Viral Diagnosis: An Understated but Essential Step for the Successful Establishment of a Diagnosis Network |
| title_full | Impact of RNA Degradation on Viral Diagnosis: An Understated but Essential Step for the Successful Establishment of a Diagnosis Network |
| title_fullStr | Impact of RNA Degradation on Viral Diagnosis: An Understated but Essential Step for the Successful Establishment of a Diagnosis Network |
| title_full_unstemmed | Impact of RNA Degradation on Viral Diagnosis: An Understated but Essential Step for the Successful Establishment of a Diagnosis Network |
| title_short | Impact of RNA Degradation on Viral Diagnosis: An Understated but Essential Step for the Successful Establishment of a Diagnosis Network |
| title_sort | impact of rna degradation on viral diagnosis an understated but essential step for the successful establishment of a diagnosis network |
| topic | viral diagnosis RNA storage secondary structures laboratory networks sample exchange real time RT-PCR |
| url | http://www.mdpi.com/2306-7381/5/1/19 |
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