Table 1: Comparison of commonly used serological assays for the diagnosis of JEV serocomplex flaviviral infections.

AssayDifferentiation between IgM and IgGTime to run assayHandling of live virus by the end userSpecies specific?#Sensitivity*Specificity*High throughputTarget flaviviral protein for serum antibodiesCommercialised or available from government Agencies?

PRNTNo, but an increased titre using paired sera is indicative of a recent infectionDaysYesNoHigh.
It is the gold standard assay
High.
It is the gold standard assay, but requires the simultaneous testing of endemic flaviviruses
NoNeutralising epitopes of the E proteinNo

HINo, but an increased titre using paired sera is indicative of a recent infectionHoursNo once the antigen has been preparedNoHighLow. Requires the simultaneous testing of endemic flavivirusesNoE proteinNo

IFAYesHoursNo once the slides have been preparedYesModerate, for example, IgM IFA not as sensitive as ELISA [128, 129].IgM: Moderate, for example, cross-reactivity 17% [130]; 14% [128]
IgG: Low, for example, cross-reactivity: 63% [130]; 45% [128]
NoAllYes, for example, Focus Diagnostics and Scimedx

ELISA IgG/IgM Yes Hours No Yes High, for example, Focus Diagnostics WNV DxSelect IgM: 93–100% [131] IgM: Moderate to High, for example, Focus Diagnostics WNV DxSelect IgM: 100% [131], but cross-reactivity with some cases of other flaviviral infections [132] Yes E protein Yes, for example, Focus Diagnostics, Panbio, CDC

Blocking/ CompetitiveNo Hours No No High, for example, [58] High, for example, [47] Yes E or NS1 No, but mAbs can be purchased from Millipore.

Immunoblot using whole viral antigenYesHoursNoYesModerate, for example, [94]Depends on the protein targeted, for example, [94]NoE, prM and NS1No

Lateral FlowIgM onlyMinutesNoYesHigh 98.8% [133]High 95.3% [133]NoE proteinYes, for example,
RapidWN [133]

MIAYesHoursNoYesHighModerate, for example, high specificity for known negative samples, but some cross-reactivity with other flaviviruses [134] High specificity for NS5 MIA [63]YesE protein [135]
NS5 [63]
Yes

#Is the assay normally set up to be specific for the species being tested, for example, horse, human, and so forth?
*The sensitivity and specificity of the assays depends on the standard to which they were compared.