4 years ago

Evaluation of a Novel Reporter Virus Neutralization Test for the Serological Diagnosis of Zika and Dengue Virus Infection.

Yang Y, Loeffelholz MJ, Kramer LD, Shi PY, Ortiz DA, Shan C, Ren P, Wong SJ
The current 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 viremia and viruria periods are brief, limiting the utility of ZIKV RNA assays. Instead, most ZIKV infections are diagnosed serologically using the 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 serologic assay was described that considerably improves the turnaround time and throughput for ZIKV diagnosis. Using the traditional PRNT as a reference method, we evaluated the performance characteristics of the reporter virus neutralization test (RVNT) on 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 a 93.1% agreement with the traditional ZIKV PRNT titers. Overall, the RVNT accurately and reliably detects neutralizing antibodies in patient serum with an improved turnaround time, and can be used for the serologic 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.

Publisher URL: https://www.ncbi.nlm.nih.gov/pubmed/28768729

DOI: PubMed:28768729

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