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Diseases of Aquatic Organisms

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DAO 117:93-106 (2015)  -  DOI: https://doi.org/10.3354/dao02937

Sturgeon nucleo-cytoplasmic large DNA virus phylogeny and PCR tests

Sharon C. Clouthier1,*, Elissa VanWalleghem1,2, Eric D. Anderson

1Fisheries & Oceans Canada, Freshwater Institute, 501 University Crescent, Winnipeg, Manitoba R3T 2N6, Canada
2University of Manitoba, Department of Biological Sciences, Winnipeg, Manitoba R3T 2N2, Canada
3Box 28, Group 30, RR2, Ste Anne, Manitoba R5H 1R2, Canada
*Corresponding author:

ABSTRACT: Sturgeon epitheliotropic nucleo-cytoplasmic large DNA viruses (NCLDVs) can cause a lethal disease of the integumentary system.  These viruses have not been assigned to any currently recognized family or genus.  In this study, phylogenetic analyses using the major capsid protein (MCP) showed that the sturgeon NCLDVs formed a cohesive taxonomic group, could be identified to the species or possibly sub-species level and formed a distinct evolutionary lineage within the Megavirales. The genetic relatedness of the sturgeon virus MCP allowed design of 3 PCR diagnostic tests with analytical specificity (ASp) inclusive of this group of viruses. The conventional PCR test, C1, had broader ASp than the 2 quantitative PCR tests, Q1 and Q2, and was inclusive of the sturgeon viruses as well as some viruses belonging to the families Mimi-, Phycodna-, or Iridoviridae. Q2 had broader specificity than Q1 but both tests recognized the sturgeon NCLDVs and did not cross-react with co-localizing sturgeon herpesviruses. Analytical test performance characteristics evaluated for Q1 and Q2 revealed sensitive assays with observed 50% limits of detection between 3 and 6.25 plasmid copies and high intra- and inter-assay repeatability. Q1 was used to test for sturgeon viruses in endangered populations of lake sturgeon Acipenser fulvescens within the Winnipeg River or Nelson River drainage systems of Manitoba, Canada. Test results indicated that namao virus is endemic in the Nelson River water basin. These tests meet the analytical requirements for diagnostic testing in Canada and are useful tools for disease management in sturgeon conservation stocking programs in North America.

KEY WORDS: Acipenseridae · Nucleo-cytoplasmic large DNA viruses · NCLDVs · Quantitative PCR · Namao virus · Mimiviridae · Endangered species

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Cite this article as: Clouthier SC, VanWalleghem E, Anderson ED (2015) Sturgeon nucleo-cytoplasmic large DNA virus phylogeny and PCR tests. Dis Aquat Org 117:93-106. https://doi.org/10.3354/dao02937

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