Diagnostic validation of three test methods for detection of cyprinid herpesvirus 3 (CyHV-3)

被引:6
作者
Clouthier, Sharon C. [1 ]
McClure, Carol [2 ]
Schroeder, Tamara [1 ]
Desai, Megan [1 ]
Hawley, Laura [3 ]
Khatkar, Sunita [1 ]
Lindsay, Melissa [1 ]
Lowe, Geoff [3 ]
Richard, Jon [3 ]
Anderson, Eric D. [4 ]
机构
[1] Fisheries & Oceans Canada, Inst Freshwater, 501 Univ Crescent, Winnipeg, MB R3T 2N6, Canada
[2] AquaEpi Res, RR3,300 Heartz Rd, Charlottetown, PE C1C 0H8, Canada
[3] Fisheries & Oceans Canada, Pacific Biol Stn, 3190 Hammond Bay Rd, Nanaimo, BC V9T 6N7, Canada
[4] Box 28,Grp 30,RR2, Ste Anne, MB R5H 1R2, Canada
关键词
CyHV-3; Diagnostic validation; Precision; Accuracy; Quantitative PCR; Virus isolation; SALMON-ANEMIA-VIRUS; EPC CELL-LINE; KOI HERPESVIRUS; THYMIDINE KINASE; PHYLOGENETIC-RELATIONSHIPS; MASS MORTALITY; TEST ACCURACY; COMMON CARP; HOT-SPOTS; DISEASE;
D O I
10.3354/dao03093
中图分类号
S9 [水产、渔业];
学科分类号
0908 ;
摘要
Cyprinid herpesvirus 3 (CyHV-3) is the aetiological agent of koi herpesvirus disease in koi and common carp. The disease is notifiable to the World Organisation for Animal Health. Three tests-quantitative polymerase chain reaction (qPCR), conventional PCR (cPCR) and virus isolation by cell culture (VI)-were validated to assess their fitness as diagnostic tools for detection of CyHV-3. Test performance metrics of diagnostic accuracy were sensitivity (DSe) and specificity (DSp). Repeatability and reproducibility were measured to assess diagnostic precision. Estimates of test accuracy, in the absence of a gold standard reference test, were generated using latent class models. Test samples originated from wild common carp naturally exposed to CyHV-3 or domesticated koi either virus free or experimentally infected with the virus. Three laboratories in Canada participated in the precision study. Moderate to high repeatability (81 to 99%) and reproducibility (72 to 97%) were observed for the qPCR and cPCR tests. The lack of agreement observed between some of the PCR test pair results was attributed to cross-contamination of samples with CyHV-3 nucleic acid. Accuracy estimates for the PCR tests were 99% for DSe and 93% for DSp. Poor precision was observed for the VI test (4 to 95%). Accuracy estimates for VI/qPCR were 90% for DSe and 88% for DSp. Collectively, the results show that the CyHV-3 qPCR test is a suitable tool for surveillance, presumptive diagnosis and certification of individuals or populations as CyHV-3 free.
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页码:101 / 122
页数:22
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