A Rapid Zika Diagnostic Assay to Measure Neutralizing Antibodies in Patients

• Published in: EBioMedicine 2017-03, Vol.17 (C), p.157-162
• Main Authors: Shan, Chao, Xie, Xuping, Ren, Ping, Loeffelholz, Michael J., Yang, Yujiao, Furuya, Andrea, Dupuis, Alan P., Kramer, Laura D., Wong, Susan J., Shi, Pei-Yong
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Neutralizing - immunology; Cell Line; Cercopithecus aethiops; Cricetinae; Cricetulus; Dengue virus; Flavivirus diagnosis; Humans; Neutralization assay; Neutralization Tests - methods; Neutralization Tests - standards; Reagent Kits, Diagnostic - standards; Research Paper; Sensitivity and Specificity; Serologic assay; Vero Cells; Zika virus; Zika Virus Infection - blood; Zika Virus Infection - immunology
• ISSN: 2352-3964; 2352-3964
• DOI: 10.1016/j.ebiom.2017.03.006

The potential association of microcephaly and other congenital abnormalities with Zika virus (ZIKV) infection during pregnancy underlines the critical need for a rapid and accurate diagnosis. Due to the short duration of ZIKV viremia in infected patients, a serologic assay that detects antibody responses to viral infection plays an essential role in diagnosing patient specimens. The current serologic diagnosis of ZIKV infection relies heavily on the labor-intensive Plaque Reduction Neutralization Test (PRNT) that requires more than one-week turnaround time and represents a major bottleneck for patient diagnosis. To overcome this limitation, we have developed a high-throughput assay for ZIKV and dengue virus (DENV) diagnosis that can attain the “gold standard” of the current PRNT assay. The new assay is homogeneous and utilizes luciferase viruses to quantify the neutralizing antibody titers in a 96-well format. Using 91 human specimens, we showed that the reporter diagnostic assay has a higher dynamic range and maintains the relative specificity of the traditional PRNT assay. Besides the improvement of assay throughput, the reporter virus technology has also shortened the turnaround time to less than two days. Collectively, our results suggest that, along with the viral RT-PCR assay, the reporter virus-based serologic assay could be potentially used as the first-line test for clinical diagnosis of ZIKV infection as well as for vaccine clinical trials. •A high-throughput assay has been developed to measure neutralizing titers of patient specimens using a reporter ZIKV.•The new assay matches the “gold standard” of the traditional labor-intensive plaque assay when testing patient specimens.•The new assay has increased the diagnostic dynamic range and shortened the turnaround time to