Development and validation of a real-time PCR assay for the detection of anguillid herpesvirus 1

S.J. van Beurden, H.A. Voorbergen-Laarman, R.E.M. Roozenburg-Hengst, Jurjen van Tellingen, O.L.M. Haenen, M.Y. Engelsma

Research output: Contribution to journalArticleAcademicpeer-review

5 Citations (Scopus)

Abstract

Anguillid herpesvirus 1 (AngHV1) causes a haemorrhagic disease with increased mortality in wild and farmed European eel, Anguilla anguilla (L.) and Japanese eel Anguilla japonica, Temminck & Schlegel). Detection of AngHV1 is currently based on virus isolation in cell culture, antibody-based typing assays or conventional PCR. We developed, optimized and concisely validated a diagnostic TaqMan probe based real-time PCR assay for the detection of AngHV1. The primers and probe target AngHV1 open reading frame 57, encoding the capsid protease and scaffold protein. Compared to conventional PCR, the developed real-time PCR is faster, less labour-intensive and has a reduced risk of cross-contamination. The real-time PCR assay was shown to be analytically sensitive and specific and has a high repeatability, efficiency and r2-value. The diagnostic performance of the assay was determined by testing 10% w/v organ suspensions and virus cultures from wild and farmed European eels from the Netherlands by conventional and real-time PCR. The developed real-time PCR assay is a useful tool for the rapid and sensitive detection of AngHV1 in 10% w/v organ suspensions from wild and farmed European eels.
Original languageEnglish
Pages (from-to)95-104
JournalJournal of Fish Diseases
Volume39
Issue number1
DOIs
Publication statusPublished - Jan 2016

Fingerprint

Herpesviridae
Eels
Anguilla anguilla
Real-Time Polymerase Chain Reaction
Anguilla
eel
quantitative polymerase chain reaction
assay
assays
Anguilla japonica
Suspensions
virus
probe
scaffolding proteins
Viruses
Polymerase Chain Reaction
capsid
cross contamination
Capsid
Netherlands

Keywords

  • AngHV1;Anguillid herpesvirus 1 ;eel herpesvirus;

Cite this

van Beurden, S.J. ; Voorbergen-Laarman, H.A. ; Roozenburg-Hengst, R.E.M. ; van Tellingen, Jurjen ; Haenen, O.L.M. ; Engelsma, M.Y. / Development and validation of a real-time PCR assay for the detection of anguillid herpesvirus 1. In: Journal of Fish Diseases. 2016 ; Vol. 39, No. 1. pp. 95-104.
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abstract = "Anguillid herpesvirus 1 (AngHV1) causes a haemorrhagic disease with increased mortality in wild and farmed European eel, Anguilla anguilla (L.) and Japanese eel Anguilla japonica, Temminck & Schlegel). Detection of AngHV1 is currently based on virus isolation in cell culture, antibody-based typing assays or conventional PCR. We developed, optimized and concisely validated a diagnostic TaqMan probe based real-time PCR assay for the detection of AngHV1. The primers and probe target AngHV1 open reading frame 57, encoding the capsid protease and scaffold protein. Compared to conventional PCR, the developed real-time PCR is faster, less labour-intensive and has a reduced risk of cross-contamination. The real-time PCR assay was shown to be analytically sensitive and specific and has a high repeatability, efficiency and r2-value. The diagnostic performance of the assay was determined by testing 10{\%} w/v organ suspensions and virus cultures from wild and farmed European eels from the Netherlands by conventional and real-time PCR. The developed real-time PCR assay is a useful tool for the rapid and sensitive detection of AngHV1 in 10{\%} w/v organ suspensions from wild and farmed European eels.",
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van Beurden, SJ, Voorbergen-Laarman, HA, Roozenburg-Hengst, REM, van Tellingen, J, Haenen, OLM & Engelsma, MY 2016, 'Development and validation of a real-time PCR assay for the detection of anguillid herpesvirus 1' Journal of Fish Diseases, vol. 39, no. 1, pp. 95-104. https://doi.org/10.1111/jfd.12330

Development and validation of a real-time PCR assay for the detection of anguillid herpesvirus 1. / van Beurden, S.J.; Voorbergen-Laarman, H.A.; Roozenburg-Hengst, R.E.M.; van Tellingen, Jurjen; Haenen, O.L.M.; Engelsma, M.Y.

In: Journal of Fish Diseases, Vol. 39, No. 1, 01.2016, p. 95-104.

Research output: Contribution to journalArticleAcademicpeer-review

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AU - van Beurden, S.J.

AU - Voorbergen-Laarman, H.A.

AU - Roozenburg-Hengst, R.E.M.

AU - van Tellingen, Jurjen

AU - Haenen, O.L.M.

AU - Engelsma, M.Y.

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AB - Anguillid herpesvirus 1 (AngHV1) causes a haemorrhagic disease with increased mortality in wild and farmed European eel, Anguilla anguilla (L.) and Japanese eel Anguilla japonica, Temminck & Schlegel). Detection of AngHV1 is currently based on virus isolation in cell culture, antibody-based typing assays or conventional PCR. We developed, optimized and concisely validated a diagnostic TaqMan probe based real-time PCR assay for the detection of AngHV1. The primers and probe target AngHV1 open reading frame 57, encoding the capsid protease and scaffold protein. Compared to conventional PCR, the developed real-time PCR is faster, less labour-intensive and has a reduced risk of cross-contamination. The real-time PCR assay was shown to be analytically sensitive and specific and has a high repeatability, efficiency and r2-value. The diagnostic performance of the assay was determined by testing 10% w/v organ suspensions and virus cultures from wild and farmed European eels from the Netherlands by conventional and real-time PCR. The developed real-time PCR assay is a useful tool for the rapid and sensitive detection of AngHV1 in 10% w/v organ suspensions from wild and farmed European eels.

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SP - 95

EP - 104

JO - Journal of Fish Diseases

JF - Journal of Fish Diseases

SN - 0140-7775

IS - 1

ER -