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The kinetics and specificity of antibody responses against Zika virus (ZIKV) are not well-characterized. This is due, in part, to the antigenic similarity between ZIKV and closely related Dengue virus (DENV) serotypes. Since these and other similar viruses co-circulate and are spread by the same mosquito species, it is difficult to disentangle ZIKV-specific antibody responses from responses to closely-related arboviruses in humans. Here we use high-density peptide microarrays to differentiate ZIKV from DENV antibody reactivity in pregnant and non-pregnant macaque monkeys with known exposure histories. We independently confirm a ZIKV-specific IgG antibody response targeting ZIKV nonstructural protein 2B (NS2B) that was recently reported in ZIKV-infected people and show that antibody reactivity in pregnant animals can be detectable as late as 113 days post-infection (dpi). We also show, however, these responses wane over time, and in one case the response was elicited following DENV infection in a previously ZIKV-exposed animal. The generally weak and short-lived antibody responses to this NS2B epitope, the only reliably ZIKV-specific epitope identified to date to our knowledge, suggest epidemiologic studies assessing seroprevalence specifically to ZIKV using linear epitope-based strategies may be prone to false negative or otherwise confounded results. These results additionally indicate some antibodies produced in response to ZIKV infection are in circulation for only a short period of time.

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