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. 2017 Aug 8;15(8):e04955. doi: 10.2903/j.efsa.2017.4955

Table 7.

Diagnostic tests for WNV

Test Target Se Sp Matrix Reference Notes
NS1‐antigen protein microarray Antibodies 95% 100% Serum Cleton et al. (in press) Differential diagnosis of flavivirus infections in horses
Real‐time RT‐PCR Antigen From 1.5 to 15 copies per reaction 100% Viral strains, human samples (cerebrospinal fluid, biopsies, serum and plasma) and mosquito pools Vázquez et al. (2016) Specificity evaluated using viral RNA from a panel of different flaviviruses and other encephalitic viruses belonging to several viral families
Real‐time RT‐PCR Antigen 80 genome copies 100% Viral strains Lineages 1 and 2 Faggioni et al. (2014) Specificity evaluated using TBE, Usutu, Dengue 1, Dengue 4, YF, JEV
SYBR Green I‐based real‐time RT‐PCR Antigen 20 copies 100% Human serum/plasma Kumar et al. (2014) Specificity evaluated using DEN‐1–4, JEV, YFV, SLEV
Antigen capture ELISA Antigen 90% 98% Human serum Saxena et al. (2013) Detection of NS1 antigen
Real‐time RT‐PCR Antigen 10 copies 100% Viral strains Barros et al. (2013) Detection and differentiation between WNV and JEV; specificity evaluated using DEN‐1–4, JEV, YFV, ZIKAV, Ntaya, TBEV, USUV, Toscana, CHIKV
Real‐time RT‐PCR Antigen 1.26 TCID50/ml for WNV‐L1, 6.3 TCID50/ml for WNV‐L2 100% Tissue, feathers, oropharyngeal and cloacal swabs and blood from wild birds, samples from mice infected experimentally Del Amo et al. (2013) Detection and differentiation between WNV and USUV; specificity evaluated using SLEV, MVEV, JEV, BAGV, DEN‐1, TBEV, VEEV, VSV, AIV, EIV, NDV, AHS4
Competitive ELISA Antibodies 100% Wild birds: 79.5% compared to VNT Sera from mammals and wild birds Sotelo et al. (2011a)
Horses: 96.5% compared to VNT
South african mammals: 79.5% compared to HAI
Giraffes: 67% compared to HAI
IgM capture ELISA Antibodies 91.7% 99.2% Horse sera Long et al. (2006)
Real‐time RT‐PCR Antigen 2–4 genome copies of WNV 100% Viral strains Eiden et al. (2010) In OIE manual. For simultaneous detection and differentiation of WNV Lineage 1 and Lineage 2. Specificity evaluated using TBEV, YFV, JEV
Nested RT‐PCR Antigen 10–8.0/100 μL ND Equine brain, blood, and cerebrospinal fluid; avian brain tissues Johnson et al. (2001) In OIE manual
Real‐time RT‐PCR Antigen 0.1 PFU 100% Human serum, CSF, brain tissue, mosquito pools, and avian tissues Lanciotti et al. (2000) In OIE manual. Specificity evaluated using DEN‐2, JEV, YFV, SLEV, Lacrosse virus, Powassan virus, MVE, WEEV, EEEV

RT‐PCR: reverse transcriptase polymerase chain reaction; IgM: immunoglobulin M; ELISA: enzyme linked immunosorbent assay; PFU: plaque‐forming unit.