Detection and surveillance of viral hemorrhagic septicemia virus using real-time RT-PCR. II. Diagnostic evaluation of two protocols

Janet V. Warg; Travis Clement; Emily R. Cornwell; Angela Cruz; Rodman G. Getchell; Cem Giray; Andrew E. Goodwin; Geoffrey H. Groocock; Mohamed Faisal; Robert Kim; Gwenn E. Merry; Nicholas B. D. Phelps; Monica M. Reising; Isaac Standish; Yan Zhang; Kathy Toohey-Kurth

doi:10.3354/dao02758

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Diseases of Aquatic Organisms

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DAO 111:15-22 (2014) - DOI: https://doi.org/10.3354/dao02758

Detection and surveillance of viral hemorrhagic septicemia virus using real-time RT-PCR. II. Diagnostic evaluation of two protocols

Janet V. Warg^1,*, Travis Clement², Emily R. Cornwell³, Angela Cruz¹, Rodman G. Getchell³, Cem Giray⁴, Andrew E. Goodwin⁵, Geoffrey H. Groocock³, Mohamed Faisal⁶, Robert Kim⁶, Gwenn E. Merry⁵, Nicholas B. D. Phelps⁷, Monica M. Reising⁸, Isaac Standish⁶, Yan Zhang⁹, Kathy Toohey-Kurth¹⁰

¹Diagnostic Virology Laboratory, National Veterinary Services Laboratories, VS, APHIS, USDA, Ames, Iowa 50010, USA
²Animal Disease Research and Diagnostic Laboratory, South Dakota State University, Brookings, South Dakota 57006, USA
³Aquatic Animal Health Program, Department of Microbiology and Immunology, Cornell University, Ithaca, New York 14853, USA
⁴Kennebec River Biosciences, Richmond, Maine 04357, USA
⁵Aquaculture/Fisheries Center, University of Arkansas Pine Bluff, Pine Bluff, Arkansas 71601, USA
⁶Aquatic Animal Health Laboratory, College of Veterinary Medicine, Michigan State University, East Lansing, Michigan 48824, USA
⁷Veterinary Diagnostic Laboratory, University of Minnesota, St. Paul, Minnesota 55108, USA
⁸Center for Veterinary Biologics, VS, APHIS, USDA, Ames, Iowa 50010, USA
⁹Animal Disease Diagnostic Laboratory, Ohio Department of Agriculture, Reynoldsburg, Ohio 43068, USA
¹⁰Wisconsin Veterinary Diagnostic Laboratory, University of Wisconsin, Madison, Wisconsin 53706, USA

*Corresponding author: janet.v.warg@aphis.usda.gov

ABSTRACT: Two real-time reverse transcription polymerase chain reaction (rRT-PCR) assays under consideration for deployment to multiple testing laboratories across the USA were evaluated for diagnostic sensitivity and specificity on tissue homogenates obtained from natural and experimental viral hemorrhagic septicemia (VHS)-infected fish. Estimates for diagnostic specificity using virus isolation as the reference method were similar between laboratories regardless of the assay. Diagnostic sensitivity estimates of 0.96 (95% CI: 0.95, 0.97) for Jonstrup et al. (2013)’s assay (J Fish Dis 36:9-23) exceeded the diagnostic sensitivity of 0.85 (95% CI: 0.83, 0.87) for Phelps et al. (2012)’s assay (J Aquat Anim Health 24:238-243). The Jonstrup rRT-PCR assay is robust as demonstrated by high sensitivity and specificity estimates across laboratories and can be used as a valuable tool for targeted surveillance and for testing of suspect VHSV samples.

KEY WORDS: VHSV · Surveillance · Real-time RT-PCR · Diagnostic sensitivity · Diagnostic specificity

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Cite this article as: Warg JV, Clement T, Cornwell ER, Cruz A and others (2014) Detection and surveillance of viral hemorrhagic septicemia virus using real-time RT-PCR. II. Diagnostic evaluation of two protocols. Dis Aquat Org 111:15-22. https://doi.org/10.3354/dao02758

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DAO 111:15-22 (2014) - DOI: https://doi.org/10.3354/dao02758

Detection and surveillance of viral hemorrhagic septicemia virus using real-time RT-PCR. II. Diagnostic evaluation of two protocols

Janet V. Warg1,*, Travis Clement2, Emily R. Cornwell3, Angela Cruz1, Rodman G. Getchell3, Cem Giray4, Andrew E. Goodwin5, Geoffrey H. Groocock3, Mohamed Faisal6, Robert Kim6, Gwenn E. Merry5, Nicholas B. D. Phelps7, Monica M. Reising8, Isaac Standish6, Yan Zhang9, Kathy Toohey-Kurth10