Development and application of a duplex PCR assay for detection of Crangon crangon bacilliform virus in populations of European brown shrimp (Crangon crangon)

Benigna Van Eynde, Olivier Christiaens, Daan Delbare, Kris Cooreman, Kelly S. Bateman, Grant D. Stentiford, Annette M. Dullemans, Monique M. van Oers, Guy Smagghe*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

1 Citation (Scopus)

Abstract

Crangon crangon bacilliform virus (CcBV) was first discovered in 2004 in European brown shrimp (Crangon crangon) caught along the English coast. This study describes a duplex PCR assay developed for the detection of CcBV, based on amplification of the lef-8 gene (211 bp) of CcBV and the E75 gene (105 bp) of C. crangon as an internal amplification control. The lef-8 and E75 primer pairs were designed based on preliminary genome sequencing information of the virus and transcriptomic data available for C. crangon, respectively. Sequencing of the resulting amplicons confirmed the specificity of this PCR assay and sequence analysis of the lef-8 fragment revealed amino acid identity percentages ranging between 31 and 42% with members of the Nudiviridae, proposing that CcBV may reside within this family.Finally, the duplex PCR assay was applied to samples of C. crangon hepatopancreas tissue collected along the Belgian coast to screen for the presence of CcBV. The prevalence of CcBV averaged 87%, which is comparable to previous reports of high prevalence, based upon histological analysis, in shrimp collected along the English coast. Development of a specific and sensitive PCR assay to detect CcBV will provide a useful tool for future aquaculture and research programs involving C. crangon.
Original languageEnglish
Pages (from-to)195-202
JournalJournal of Invertebrate Pathology
Volume153
Early online date14 Mar 2018
DOIs
Publication statusPublished - Mar 2018

Fingerprint

Crangon crangon
duplex
virus
shrimp
assay
viruses
assays
coast
amplification
coasts
detection
gene
research program
aquaculture
genome
amino acid
hepatopancreas
research programs
transcriptomics

Keywords

  • Aquaculture
  • CcBV
  • Crustacean
  • Diagnostic
  • Disease
  • Nudiviridae

Cite this

Van Eynde, Benigna ; Christiaens, Olivier ; Delbare, Daan ; Cooreman, Kris ; Bateman, Kelly S. ; Stentiford, Grant D. ; Dullemans, Annette M. ; van Oers, Monique M. ; Smagghe, Guy. / Development and application of a duplex PCR assay for detection of Crangon crangon bacilliform virus in populations of European brown shrimp (Crangon crangon). In: Journal of Invertebrate Pathology. 2018 ; Vol. 153. pp. 195-202.
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abstract = "Crangon crangon bacilliform virus (CcBV) was first discovered in 2004 in European brown shrimp (Crangon crangon) caught along the English coast. This study describes a duplex PCR assay developed for the detection of CcBV, based on amplification of the lef-8 gene (211 bp) of CcBV and the E75 gene (105 bp) of C. crangon as an internal amplification control. The lef-8 and E75 primer pairs were designed based on preliminary genome sequencing information of the virus and transcriptomic data available for C. crangon, respectively. Sequencing of the resulting amplicons confirmed the specificity of this PCR assay and sequence analysis of the lef-8 fragment revealed amino acid identity percentages ranging between 31 and 42{\%} with members of the Nudiviridae, proposing that CcBV may reside within this family.Finally, the duplex PCR assay was applied to samples of C. crangon hepatopancreas tissue collected along the Belgian coast to screen for the presence of CcBV. The prevalence of CcBV averaged 87{\%}, which is comparable to previous reports of high prevalence, based upon histological analysis, in shrimp collected along the English coast. Development of a specific and sensitive PCR assay to detect CcBV will provide a useful tool for future aquaculture and research programs involving C. crangon.",
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Development and application of a duplex PCR assay for detection of Crangon crangon bacilliform virus in populations of European brown shrimp (Crangon crangon). / Van Eynde, Benigna; Christiaens, Olivier; Delbare, Daan; Cooreman, Kris; Bateman, Kelly S.; Stentiford, Grant D.; Dullemans, Annette M.; van Oers, Monique M.; Smagghe, Guy.

In: Journal of Invertebrate Pathology, Vol. 153, 03.2018, p. 195-202.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Development and application of a duplex PCR assay for detection of Crangon crangon bacilliform virus in populations of European brown shrimp (Crangon crangon)

AU - Van Eynde, Benigna

AU - Christiaens, Olivier

AU - Delbare, Daan

AU - Cooreman, Kris

AU - Bateman, Kelly S.

AU - Stentiford, Grant D.

AU - Dullemans, Annette M.

AU - van Oers, Monique M.

AU - Smagghe, Guy

PY - 2018/3

Y1 - 2018/3

N2 - Crangon crangon bacilliform virus (CcBV) was first discovered in 2004 in European brown shrimp (Crangon crangon) caught along the English coast. This study describes a duplex PCR assay developed for the detection of CcBV, based on amplification of the lef-8 gene (211 bp) of CcBV and the E75 gene (105 bp) of C. crangon as an internal amplification control. The lef-8 and E75 primer pairs were designed based on preliminary genome sequencing information of the virus and transcriptomic data available for C. crangon, respectively. Sequencing of the resulting amplicons confirmed the specificity of this PCR assay and sequence analysis of the lef-8 fragment revealed amino acid identity percentages ranging between 31 and 42% with members of the Nudiviridae, proposing that CcBV may reside within this family.Finally, the duplex PCR assay was applied to samples of C. crangon hepatopancreas tissue collected along the Belgian coast to screen for the presence of CcBV. The prevalence of CcBV averaged 87%, which is comparable to previous reports of high prevalence, based upon histological analysis, in shrimp collected along the English coast. Development of a specific and sensitive PCR assay to detect CcBV will provide a useful tool for future aquaculture and research programs involving C. crangon.

AB - Crangon crangon bacilliform virus (CcBV) was first discovered in 2004 in European brown shrimp (Crangon crangon) caught along the English coast. This study describes a duplex PCR assay developed for the detection of CcBV, based on amplification of the lef-8 gene (211 bp) of CcBV and the E75 gene (105 bp) of C. crangon as an internal amplification control. The lef-8 and E75 primer pairs were designed based on preliminary genome sequencing information of the virus and transcriptomic data available for C. crangon, respectively. Sequencing of the resulting amplicons confirmed the specificity of this PCR assay and sequence analysis of the lef-8 fragment revealed amino acid identity percentages ranging between 31 and 42% with members of the Nudiviridae, proposing that CcBV may reside within this family.Finally, the duplex PCR assay was applied to samples of C. crangon hepatopancreas tissue collected along the Belgian coast to screen for the presence of CcBV. The prevalence of CcBV averaged 87%, which is comparable to previous reports of high prevalence, based upon histological analysis, in shrimp collected along the English coast. Development of a specific and sensitive PCR assay to detect CcBV will provide a useful tool for future aquaculture and research programs involving C. crangon.

KW - Aquaculture

KW - CcBV

KW - Crustacean

KW - Diagnostic

KW - Disease

KW - Nudiviridae

U2 - 10.1016/j.jip.2018.03.006

DO - 10.1016/j.jip.2018.03.006

M3 - Article

VL - 153

SP - 195

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JO - Journal of Invertebrate Pathology

JF - Journal of Invertebrate Pathology

SN - 0022-2011

ER -