A High-Performance Multiplex Immunoassay for Serodiagnosis of Flavivirus-Associated Neurological Diseases in Horses

Table 3

Comparison of the earliest antibody detection using flavivirus ELISA, MIA, and VNT. Identification of specific WNV, JEV, or TBEV antibodies in reference sera sampled on days 0, 4, 8, 11, 14, 20, 35, and 58.

		ELISA positive^(1)	MIA		MNT
			Positive for WNV.sE^(2)	Flavivirus; identification^(3)	Positive^(4)	Flavivirus identification^(5)
Sera	WNV; lineage 1	D11	D11	D35	D8	D14
	WNV; lineage 2	D11	D8	D8	D8	D14
	JEV; TBEV	D20; D20	D20; D20	Not achieved; D35	D20; D8	D35; D8

⁽¹⁾ID Screen West Nile competition ELISA kit (ID Vet). Sample positive if %S/N 40%, doubtful if 40 %S/N 50%, and negative if %S/N 50.
⁽²⁾WNV.sE positive if S/P ratio 17 for the WNV.sE bead.
⁽³⁾WNV.EDIII identification if S/P ratio 17 for the WNV.sE bead and S/P ratio > 54 for the WNV.EDIII bead.
JEV.EDIII identification if S/P ratio > 17 for the WNV.sE bead and S/P ratio > 55 for the JEV.EDIII bead; TBEV.EDIII identification if S/P ratio 61 for the TBEV.EDIII bead.
⁽⁴⁾MNT positive if MNT titre threshold (10 for JEV and WNV and 20 for TBEV).
⁽⁵⁾MNT identification of the flavivirus with the highest neutralisation capacity and at least a fourfold difference in titres.