Rapid Molecular Detection of Zika Virus in Acute-Phase Urine Samples Using the Recombinase Polymerase Amplification Assay

PLoS Curr. 2017 Jan 25:9:ecurrents.outbreaks.a7f1db2c7d66c3fc0ea0a774305d319e. doi: 10.1371/currents.outbreaks.a7f1db2c7d66c3fc0ea0a774305d319e.

Abstract

Background: Currently the detection of Zika virus (ZIKV) in patient samples is done by real-time RT-PCR. Samples collected from rural area are sent to highly equipped laboratories for screening. A rapid point-of-care test is needed to detect the virus, especially at low resource settings.

Methodology/principal findings: In this report, we describe the development of a reverse transcription isothermal recombinase polymerase amplification (RT-RPA) assay for the identification of ZIKV. RT-RPA assay was portable, sensitive (21 RNA molecules), and rapid (3-15 minutes). No cross-reactivity was detected to other flaviviruses, alphaviruses and arboviruses. Compared to real-time RT-PCR, the diagnostic sensitivity was 92%, while the specificity was 100%.

Conclusions/significance: The developed assay is a promising platform for rapid point of need detection of ZIKV in low resource settings and elsewhere (e.g. during mass gathering).

Keywords: diagnostics; point of need; zika virus.

Grants and funding

Universitätsbund Göttingen e.V. provided Ahmed Abd El Wahed with travel expenses to support the field trial in Brazil (ID: VAB6/16WN3). The funders had no role in design of the study, data collection and analysis, decision to publish, or preparation of the manuscript.