Detection of Infectious Pancreatic Necrosis Virus in Subclinically Infected Atlantic Salmon by Virus Isolation in Cell Culture or Real-Time Reverse Transcription Polymerase Chain Reaction: Influence of Sample Preservation and Storage
Infectious pancreatic necrosis, an important problem of the salmon industry worldwide, is caused by Infectious pancreatic necrosis virus (IPNV). Fish surviving an IPNV infection become virus carriers, and the identification of infected fish is highly relevant to disease control. The target organ for...
Published in: | Journal of Veterinary Diagnostic Investigation |
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crsagepubl:10.1177/104063871002200606 2024-09-15T17:56:07+00:00 Detection of Infectious Pancreatic Necrosis Virus in Subclinically Infected Atlantic Salmon by Virus Isolation in Cell Culture or Real-Time Reverse Transcription Polymerase Chain Reaction: Influence of Sample Preservation and Storage Ørpetveit, Irene Mikalsen, Aase B. Sindre, Hilde Evensen, Øystein Dannevig, Birgit H. Midtlyng, Paul J. 2010 http://dx.doi.org/10.1177/104063871002200606 http://journals.sagepub.com/doi/pdf/10.1177/104063871002200606 http://journals.sagepub.com/doi/full-xml/10.1177/104063871002200606 en eng SAGE Publications http://journals.sagepub.com/page/policies/text-and-data-mining-license Journal of Veterinary Diagnostic Investigation volume 22, issue 6, page 886-895 ISSN 1040-6387 1943-4936 journal-article 2010 crsagepubl https://doi.org/10.1177/104063871002200606 2024-09-03T04:20:14Z Infectious pancreatic necrosis, an important problem of the salmon industry worldwide, is caused by Infectious pancreatic necrosis virus (IPNV). Fish surviving an IPNV infection become virus carriers, and the identification of infected fish is highly relevant to disease control. The target organ for IPNV diagnosis is the kidney, where the virus persists, usually with low virus loads. The current study documents a real-time reverse transcription polymerase chain reaction (real-time RT-PCR) assay that proved 100 times more sensitive than a conventional RT-PCR. Cell culture and real-time RT-PCR were compared for their ability to detect IPNV in carrier Atlantic salmon kidney samples after different preservation and storage procedures. Storage of whole tissue at −80°C for 1 month and storage of tissue homogenized in transport medium (TM) at +4°C for 1 week before investigation in cell cultures resulted in a marked reduction of virus infectivity. For detection by real-time RT-PCR, storage of whole tissue was suboptimal, whereas storage of tissue homogenized in TM did not affect virus detection. The results of the present study demonstrate that both cell culture and real-time RT-PCR are reliable tests for the detection of low amounts of IPNV in kidneys of carrier Atlantic salmon, and both methods are relatively robust against minor preservation and storage deviations, or both. Preservation of tissues in RNA stabilization solution seems only necessary when samples are to be shipped at ambient temperatures or when laboratory testing might be delayed. Independent of detection method, these results indicate that for long-term storage, samples are best kept at −80°C after homogenization in TM. Article in Journal/Newspaper Atlantic salmon SAGE Publications Journal of Veterinary Diagnostic Investigation 22 6 886 895 |
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English |
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Infectious pancreatic necrosis, an important problem of the salmon industry worldwide, is caused by Infectious pancreatic necrosis virus (IPNV). Fish surviving an IPNV infection become virus carriers, and the identification of infected fish is highly relevant to disease control. The target organ for IPNV diagnosis is the kidney, where the virus persists, usually with low virus loads. The current study documents a real-time reverse transcription polymerase chain reaction (real-time RT-PCR) assay that proved 100 times more sensitive than a conventional RT-PCR. Cell culture and real-time RT-PCR were compared for their ability to detect IPNV in carrier Atlantic salmon kidney samples after different preservation and storage procedures. Storage of whole tissue at −80°C for 1 month and storage of tissue homogenized in transport medium (TM) at +4°C for 1 week before investigation in cell cultures resulted in a marked reduction of virus infectivity. For detection by real-time RT-PCR, storage of whole tissue was suboptimal, whereas storage of tissue homogenized in TM did not affect virus detection. The results of the present study demonstrate that both cell culture and real-time RT-PCR are reliable tests for the detection of low amounts of IPNV in kidneys of carrier Atlantic salmon, and both methods are relatively robust against minor preservation and storage deviations, or both. Preservation of tissues in RNA stabilization solution seems only necessary when samples are to be shipped at ambient temperatures or when laboratory testing might be delayed. Independent of detection method, these results indicate that for long-term storage, samples are best kept at −80°C after homogenization in TM. |
format |
Article in Journal/Newspaper |
author |
Ørpetveit, Irene Mikalsen, Aase B. Sindre, Hilde Evensen, Øystein Dannevig, Birgit H. Midtlyng, Paul J. |
spellingShingle |
Ørpetveit, Irene Mikalsen, Aase B. Sindre, Hilde Evensen, Øystein Dannevig, Birgit H. Midtlyng, Paul J. Detection of Infectious Pancreatic Necrosis Virus in Subclinically Infected Atlantic Salmon by Virus Isolation in Cell Culture or Real-Time Reverse Transcription Polymerase Chain Reaction: Influence of Sample Preservation and Storage |
author_facet |
Ørpetveit, Irene Mikalsen, Aase B. Sindre, Hilde Evensen, Øystein Dannevig, Birgit H. Midtlyng, Paul J. |
author_sort |
Ørpetveit, Irene |
title |
Detection of Infectious Pancreatic Necrosis Virus in Subclinically Infected Atlantic Salmon by Virus Isolation in Cell Culture or Real-Time Reverse Transcription Polymerase Chain Reaction: Influence of Sample Preservation and Storage |
title_short |
Detection of Infectious Pancreatic Necrosis Virus in Subclinically Infected Atlantic Salmon by Virus Isolation in Cell Culture or Real-Time Reverse Transcription Polymerase Chain Reaction: Influence of Sample Preservation and Storage |
title_full |
Detection of Infectious Pancreatic Necrosis Virus in Subclinically Infected Atlantic Salmon by Virus Isolation in Cell Culture or Real-Time Reverse Transcription Polymerase Chain Reaction: Influence of Sample Preservation and Storage |
title_fullStr |
Detection of Infectious Pancreatic Necrosis Virus in Subclinically Infected Atlantic Salmon by Virus Isolation in Cell Culture or Real-Time Reverse Transcription Polymerase Chain Reaction: Influence of Sample Preservation and Storage |
title_full_unstemmed |
Detection of Infectious Pancreatic Necrosis Virus in Subclinically Infected Atlantic Salmon by Virus Isolation in Cell Culture or Real-Time Reverse Transcription Polymerase Chain Reaction: Influence of Sample Preservation and Storage |
title_sort |
detection of infectious pancreatic necrosis virus in subclinically infected atlantic salmon by virus isolation in cell culture or real-time reverse transcription polymerase chain reaction: influence of sample preservation and storage |
publisher |
SAGE Publications |
publishDate |
2010 |
url |
http://dx.doi.org/10.1177/104063871002200606 http://journals.sagepub.com/doi/pdf/10.1177/104063871002200606 http://journals.sagepub.com/doi/full-xml/10.1177/104063871002200606 |
genre |
Atlantic salmon |
genre_facet |
Atlantic salmon |
op_source |
Journal of Veterinary Diagnostic Investigation volume 22, issue 6, page 886-895 ISSN 1040-6387 1943-4936 |
op_rights |
http://journals.sagepub.com/page/policies/text-and-data-mining-license |
op_doi |
https://doi.org/10.1177/104063871002200606 |
container_title |
Journal of Veterinary Diagnostic Investigation |
container_volume |
22 |
container_issue |
6 |
container_start_page |
886 |
op_container_end_page |
895 |
_version_ |
1810432322906882048 |