Currently, the laboratory diagnosis of Zika virus (
ZIKV) infection is primarily through the detection of ZIKV
RNA or
antibodies against ZIKV
proteins. The detection of
viral RNA is highly sensitive and specific, but periods of
viremia and viruria are brief, limiting the utility of ZIKV
RNA assays. Instead, most ZIKV
infections are diagnosed serologically, using an
IgM antibody capture
enzyme-linked
immunosorbent assay (MAC-ELISA) for screening, followed by a confirmatory plaque reduction neutralization test (PRNT). Typical turnaround times vary, due to assay incubation periods and a lack of clinical laboratories performing these tests. Recently, a novel
luciferase-ZIKV- and -dengue virus (DENV)-based serological assay, which considerably improves the turnaround times and throughput for ZIKV diagnosis, was described. Using the traditional PRNT as a reference method, we evaluated the performance characteristics of the reporter virus neutralization test (RVNT) with 258 clinical serum specimens. The ZIKV RVNT produced primary ZIKV screening and secondary confirmation results in 4 days, with 100% reproducibility. As a screening assay, the ZIKV RVNT displayed excellent diagnostic accuracy, sensitivity, and specificity of 98.2%, 100%, and 98.1%, respectively. As a confirmatory assay, the ZIKV RVNT titers displayed 93.1% agreement with the traditional ZIKV PRNT titers. Overall, the RVNT accurately and reliably detects
neutralizing antibodies in patient serum specimens, with improved turnaround times, and can be used for the serological detection of ZIKV
infections. Due to the homogeneous 96-well format, the RVNT has also significantly improved the assay throughput to allow testing of a large number of specimens in a single run.