Table 1.

Serum panels used for development and evaluation of the FLI HeV DIVA ELISA using HeV-G and HeV-N proteins.

Panel	Purpose, Determination of	Source of Serum	Assays Performed
1	Cut-off DSp	German negative horses (n = 288); These samples had been submitted from different clinics to the National Reference Laboratory for West Nile Virus (WNV) between 2009 and 2012 for WNV screening. None of the horses had a history of travelling to Australia or being vaccinated against HeV, and these samples were therefore considered HeV-negative	Preliminary Cut-off determination ROC curve analysis FLI HeV DIVA ELISA, ACDP HeV DIVA ELISA, HeV VNT
2	Cut-off DSp	Australian negative horses (n = 105)	Cut-off determination ROC curve analysis BLCM FLI HeV DIVA ELISA, ACDP HeV DIVA ELISA, HeV VNT
3	Cut-off DSe, DSp	Australian vaccinated horses (n = 40); diagnostic field samples	FLI HeV DIVA ELISA, ACDP HeV DIVA ELISA BLCM
4	Cut-off DSe ASe	Australian HeV-positive samples (n = 21) from outbreak episodes (QLD) and follow-up testing	Cut-off determination and ROC curve analysis BLCM FLI HeV DIVA ELISA, ACDP HeV DIVA ELISA, HeV VNT
5	ASp	Serum samples originating from different species (n = 17) including horses, guinea pigs, pigs, rabbits, and goats, containing antibodies against different paramyxoviruses (peste des petits ruminants virus, rinderpest virus, canine distemper virus, Newcastle disease virus, parainfluenzavirus type 1–4, mumps virus, Nariva virus, Tioman virus, Menangle virus, blue eye rubulavirus, Mossman virus)	FLI HeV DIVA ELISA

FLI = Friedrich Loeffler Institut; ACDP = Australian Centre for Disease Preparedness; QLD = Queensland; ASe = analytical sensitivity; ASp = analytical specificity; DSe = diagnostic sensitivity; DSp = diagnostic specificity; BLCM = Bayesian latent class model.