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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)

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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.
Keywords
Crustacean, Disease, CcBV, Nudiviridae, Diagnostic, Aquaculture, SPOT SYNDROME VIRUS, YELLOW-HEAD VIRUS, INTERNAL AMPLIFICATION CONTROLS, POLYMERASE-CHAIN-REACTION, FRESH-WATER CRAYFISH, PENAEID SHRIMP, MONOCLONAL-ANTIBODIES, PARALYSIS-VIRUS, DNA EXTRACTION, NECROSIS VIRUS

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Chicago
Van Eynde, Benigna, Olivier Christiaens, Daan Delbare, Kris Cooreman, Kelly S Bateman, Grant D Stentiford, Annette M Dullemans, Monique M van Oers, and Guy Smagghe. 2018. “Development and Application of a Duplex PCR Assay for Detection of Crangon Crangon Bacilliform Virus in Populations of European Brown Shrimp (Crangon Crangon).” Journal of Invertebrate Pathology 153: 195–202.
APA
Van Eynde, B., Christiaens, O., Delbare, D., Cooreman, K., Bateman, K. S., Stentiford, G. D., Dullemans, A. M., et al. (2018). Development and application of a duplex PCR assay for detection of Crangon crangon bacilliform virus in populations of European brown shrimp (Crangon crangon). JOURNAL OF INVERTEBRATE PATHOLOGY, 153, 195–202.
Vancouver
1.
Van Eynde B, Christiaens O, Delbare D, Cooreman K, Bateman KS, Stentiford GD, et al. Development and application of a duplex PCR assay for detection of Crangon crangon bacilliform virus in populations of European brown shrimp (Crangon crangon). JOURNAL OF INVERTEBRATE PATHOLOGY. 2018;153:195–202.
MLA
Van Eynde, Benigna, Olivier Christiaens, Daan Delbare, et al. “Development and Application of a Duplex PCR Assay for Detection of Crangon Crangon Bacilliform Virus in Populations of European Brown Shrimp (Crangon Crangon).” JOURNAL OF INVERTEBRATE PATHOLOGY 153 (2018): 195–202. Print.
@article{8557720,
  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.},
  author       = {Van Eynde, Benigna and Christiaens, Olivier and Delbare, Daan and Cooreman, Kris and Bateman, Kelly S and Stentiford, Grant D and Dullemans, Annette M and van Oers, Monique M and Smagghe, Guy},
  issn         = {0022-2011},
  journal      = {JOURNAL OF INVERTEBRATE PATHOLOGY},
  keyword      = {Crustacean,Disease,CcBV,Nudiviridae,Diagnostic,Aquaculture,SPOT SYNDROME VIRUS,YELLOW-HEAD VIRUS,INTERNAL AMPLIFICATION CONTROLS,POLYMERASE-CHAIN-REACTION,FRESH-WATER CRAYFISH,PENAEID SHRIMP,MONOCLONAL-ANTIBODIES,PARALYSIS-VIRUS,DNA EXTRACTION,NECROSIS VIRUS},
  language     = {eng},
  pages        = {195--202},
  title        = {Development and application of a duplex PCR assay for detection of Crangon crangon bacilliform virus in populations of European brown shrimp (Crangon crangon)},
  url          = {http://dx.doi.org/10.1016/j.jip.2018.03.006},
  volume       = {153},
  year         = {2018},
}

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