Diagnosis of Zika Virus Infection by Peptide Array and Enzyme-Linked Immunosorbent Assay.

Nischay Mishra,Diego A Espinosa,Thomas Briese,Adrian Caciula,Lokendra V Chauhan,Adam Price,W Ian Lipkin,Eva Harris,Richard G Jarman,Jennifer L Rakeman,Xiaoyu Che,Magelda Montoya Cruz,James Ng,Riddhi Thakkar,Rafal Tokarz,Komal Jain,Ángel Balmaseda ,Jigar Patel ,Chantal Reusken ,Christina Egan ,Marion Koopmans ,Eric Sullivan

doi:10.1128/mbio.00095-18

Abstract

ABSTRACTZika virus (ZIKV) is implicated in fetal stillbirth, microcephaly, intracranial calcifications, and ocular anomalies following vertical transmission from infected mothers. In adults, infection may trigger autoimmune inflammatory polyneuropathy. Transmission most commonly follows the bite of infected Aedes mosquitoes but may also occur through sexual intercourse or receipt of blood products. Definitive diagnosis through detection of viral RNA is possible in serum or plasma within 10 days of disease onset, in whole blood within 3 weeks of onset, and in semen for up to 3 months. Serological diagnosis is nonetheless critical because few patients have access to molecular diagnostics during the acute phase of infection and infection may be associated with only mild or inapparent disease that does not prompt molecular testing. Serological diagnosis is confounded by cross-reactivity of immune sera with other flaviviruses endemic in the areas where ZIKV has recently emerged. Accordingly, we built a high-density microarray comprising nonredundant 12-mer peptides that tile, with one-residue overlap, the proteomes of Zika, dengue, yellow fever, West Nile, Ilheus, Oropouche, and chikungunya viruses. Serological analysis enabled discovery of a ZIKV NS2B 20-residue peptide that had high sensitivity (96.0%) and specificity (95.9%) versus natural infection with or vaccination against dengue, chikungunya, yellow fever, West Nile, tick-borne encephalitis, or Japanese encephalitis virus in a microarray assay and an enzyme-linked immunosorbent assay (ELISA) of early-convalescent-phase sera (2 to 3 weeks after onset of symptomatic infection).

Highlights

Zika virus (ZIKV) is implicated in fetal stillbirth, microcephaly, intracranial calcifications, and ocular anomalies following vertical transmission from infected mothers
The 12-mer format is based on the observation that serum antibodies bind linear peptide sequences ranging from 5 to 9 amino acids and bind most efficiently when targets are flanked by additional amino acids [13]
Since the emergence of ZIKV in the Americas in 2015, 583,144 cases have been reported to the World Health Organization

Summary

Introduction

Zika virus (ZIKV) is implicated in fetal stillbirth, microcephaly, intracranial calcifications, and ocular anomalies following vertical transmission from infected mothers. Serological analysis enabled discovery of a ZIKV NS2B 20-residue peptide that had high sensitivity (96.0%) and specificity (95.9%) versus natural infection with or vaccination against dengue, chikungunya, yellow fever, West Nile, tick-borne encephalitis, or Japanese encephalitis virus in a microarray assay and an enzyme-linked immunosorbent assay (ELISA) of early-convalescent-phase sera (2 to 3 weeks after onset of symptomatic infection). Using high-density peptide arrays that tile the proteomes of a selection of flaviviruses to identify a ZIKV-specific peptide, we established two assays that enable sensitive and specific diagnosis of exposure to ZIKV. Cross-reactivity between ZIKV and other flaviviruses may complicate differential serodiagnosis and efforts to investigate the epidemiology of infection and linkage to disease This is challenging in areas where dengue virus (DENV) is endemic or Japanese encephalitis virus (JEV) or yellow fever virus (YFV) vaccination is common. We adapted this NS2B peptide to a synthetic biotinylated peptide IgG ELISA system with the goal of establishing an inexpensive, rapid, sensitive, specific, and point-of-care method for ZIKV IgG detection

Methods

Results

Conclusion