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

Research output: Contribution to journalArticlepeer-review


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.

Original languageEnglish (US)
Pages (from-to)15-22
Number of pages8
JournalDiseases of aquatic organisms
Issue number1
StatePublished - Aug 21 2014


  • Diagnostic sensitivity
  • Diagnostic specificity
  • Real-time RT-PCR
  • Surveillance
  • VHSV

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