Evaluation of a rapid method for recovery of norovirus and hepatitis A virus from oysters and blue mussels

Katrine Uhrbrand, Mette Myrmel, Leena Maunula, Kirsti Vainio, Ramona Trebbien, Birgit Nørrung, Anna Charlotte Schultz

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Foodborne outbreaks caused by noroviruses (NoVs) and hepatitis A virus (HAV) are often linked to consumption of contaminated shellfish. The objective of this study was to identify an appropriate virus recovery method for real-time reverse transcriptase (RT)-PCR detection and subsequently to evaluate this method on shellfish bioaccumulated with virus in a collaborative study. Five methods were compared for recovery of NoV GII.7 and feline calicivirus from spiked digestive tissue of oysters and mussels. A method based on proteinase K digestion followed by NucliSENS miniMAG extraction was found to be the most efficient with a 50% limit of detection (LOD50) of 62 and 12 RT-PCR U/1.5 g digestive tissue for NoV GII.7 in oysters and mussels, respectively. Evaluation of the method in four laboratories found the percentage of sensitivity, based on low/high levels of virus bioaccumulated in oysters, to be 33/80 for NoV GI.3b, 13/92 for NoV GII.4 and 50/42 for HAV. A specificity of 100% was found for all three viruses in non-bioaccumulated oysters. As process control Mengovirus (vMC0) showed an average recovery of 1.8% from oysters and 1.2% from mussels. The study demonstrates that this recovery method can be useful for harmonized data generation and routine viral analyses of shellfish.
Original languageEnglish
JournalJournal of Virological Methods
Volume169
Issue number1
Pages (from-to)70-78
ISSN0166-0934
DOIs
Publication statusPublished - 2010

Keywords

  • Norovirus
  • Hepatitis A virus
  • Oyster
  • Mussel
  • Virus recovery
  • Real-time RT-PCR detection

Cite this

Uhrbrand, Katrine ; Myrmel, Mette ; Maunula, Leena ; Vainio, Kirsti ; Trebbien, Ramona ; Nørrung, Birgit ; Schultz, Anna Charlotte. / Evaluation of a rapid method for recovery of norovirus and hepatitis A virus from oysters and blue mussels. In: Journal of Virological Methods. 2010 ; Vol. 169, No. 1. pp. 70-78.
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abstract = "Foodborne outbreaks caused by noroviruses (NoVs) and hepatitis A virus (HAV) are often linked to consumption of contaminated shellfish. The objective of this study was to identify an appropriate virus recovery method for real-time reverse transcriptase (RT)-PCR detection and subsequently to evaluate this method on shellfish bioaccumulated with virus in a collaborative study. Five methods were compared for recovery of NoV GII.7 and feline calicivirus from spiked digestive tissue of oysters and mussels. A method based on proteinase K digestion followed by NucliSENS miniMAG extraction was found to be the most efficient with a 50{\%} limit of detection (LOD50) of 62 and 12 RT-PCR U/1.5 g digestive tissue for NoV GII.7 in oysters and mussels, respectively. Evaluation of the method in four laboratories found the percentage of sensitivity, based on low/high levels of virus bioaccumulated in oysters, to be 33/80 for NoV GI.3b, 13/92 for NoV GII.4 and 50/42 for HAV. A specificity of 100{\%} was found for all three viruses in non-bioaccumulated oysters. As process control Mengovirus (vMC0) showed an average recovery of 1.8{\%} from oysters and 1.2{\%} from mussels. The study demonstrates that this recovery method can be useful for harmonized data generation and routine viral analyses of shellfish.",
keywords = "Norovirus, Hepatitis A virus, Oyster, Mussel, Virus recovery, Real-time RT-PCR detection",
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Evaluation of a rapid method for recovery of norovirus and hepatitis A virus from oysters and blue mussels. / Uhrbrand, Katrine; Myrmel, Mette; Maunula, Leena; Vainio, Kirsti; Trebbien, Ramona; Nørrung, Birgit; Schultz, Anna Charlotte.

In: Journal of Virological Methods, Vol. 169, No. 1, 2010, p. 70-78.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

T1 - Evaluation of a rapid method for recovery of norovirus and hepatitis A virus from oysters and blue mussels

AU - Uhrbrand, Katrine

AU - Myrmel, Mette

AU - Maunula, Leena

AU - Vainio, Kirsti

AU - Trebbien, Ramona

AU - Nørrung, Birgit

AU - Schultz, Anna Charlotte

PY - 2010

Y1 - 2010

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AB - Foodborne outbreaks caused by noroviruses (NoVs) and hepatitis A virus (HAV) are often linked to consumption of contaminated shellfish. The objective of this study was to identify an appropriate virus recovery method for real-time reverse transcriptase (RT)-PCR detection and subsequently to evaluate this method on shellfish bioaccumulated with virus in a collaborative study. Five methods were compared for recovery of NoV GII.7 and feline calicivirus from spiked digestive tissue of oysters and mussels. A method based on proteinase K digestion followed by NucliSENS miniMAG extraction was found to be the most efficient with a 50% limit of detection (LOD50) of 62 and 12 RT-PCR U/1.5 g digestive tissue for NoV GII.7 in oysters and mussels, respectively. Evaluation of the method in four laboratories found the percentage of sensitivity, based on low/high levels of virus bioaccumulated in oysters, to be 33/80 for NoV GI.3b, 13/92 for NoV GII.4 and 50/42 for HAV. A specificity of 100% was found for all three viruses in non-bioaccumulated oysters. As process control Mengovirus (vMC0) showed an average recovery of 1.8% from oysters and 1.2% from mussels. The study demonstrates that this recovery method can be useful for harmonized data generation and routine viral analyses of shellfish.

KW - Norovirus

KW - Hepatitis A virus

KW - Oyster

KW - Mussel

KW - Virus recovery

KW - Real-time RT-PCR detection

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DO - 10.1016/j.jviromet.2010.06.019

M3 - Journal article

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EP - 78

JO - Journal of Virological Methods

JF - Journal of Virological Methods

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ER -