Diagnosing Zika virus infection against a background of other flaviviruses: Studies in high resolution serological analysis

Sören Hansen,Ahmed Abd El Wahed,Susanne Böhlken-Fascher,Ronald Frank,Matthias Niedrig,Oumar Faye,Amadou A Sall,Hans-Joachim Fritz,Sven-Kevin Hotop,Jonas Schmidt-Chanasit,Claus-Peter Czerny,Sabri S Sanabani,Oumar Ndiaye,Frank Hufert,Rodrigo Pessôa,Christiane Stahl-Hennig,Mark Brönstrup

doi:10.1038/s41598-019-40224-2

Abstract

Zika virus (ZIKV) is a mosquito-borne flavivirus. Homologous proteins of different flaviviruses display high degrees of sequence identity, especially within subgroups. This leads to extensive immunological cross-reactivity and corresponding problems for developing a ZIKV-specific serological assay. In this study, peptide microarrays were employed to identify individual ZIKV antibody targets with promise in differential diagnosis. A total of 1643 overlapping oligopeptides were synthesized and printed onto glass slides. Together, they encompass the full amino acid sequences of ZIKV proteomes of African, Brazilian, USA, and French Polynesian origins. The resulting ZIKV scanning microarray chips were used to screen three pools of sera from recent Zika outbreaks in Senegal and Cape Verde, in Brazil, and from overseas travelers returning to the EU. Together with a mixed pool of well characterized, archived sera of patients suffering from infections by dengue, yellow fever, tick-borne encephalitis, and West Nile viruses, a total of 42 sera went into the study. Sixty-eight antibody target regions were identified. Most of which were hitherto unknown. Alignments and sequence comparisons revealed 13 of which could be classified as bona fide ZIKV-specific. These identified antibody target regions constitute a founding set of analytical tools for serological discrimination of ZIKV from other flaviviruses.

Highlights

IntroductionHomologous proteins of different flaviviruses display high degrees of sequence identity, especially within subgroups
Zika virus (ZIKV) is a mosquito-borne flavivirus
Since a spots series as such often does not allow distinguishing between single epitope and antibody target region (ATR), the latter term is occasionally used as a synonym of the generic name “antibody target” with epitopes regarded as extreme cases within that class

Summary

Introduction

Homologous proteins of different flaviviruses display high degrees of sequence identity, especially within subgroups This leads to extensive immunological cross-reactivity and corresponding problems for developing a ZIKV-specific serological assay. Alignments and sequence comparisons revealed 13 of which could be classified as bona fide ZIKV-specific These identified antibody target regions constitute a founding set of analytical tools for serological discrimination of ZIKV from other flaviviruses. High degrees of sequence identity of homologous proteins and concomitant immunological cross-reactivity among all flaviviruses, in particular within the various subgroups, have been reported[18,19] These facts profoundly complicate serological diagnosis, especially in areas, where ZIKV and dengue virus are co-circulating and/or yellow fever vaccine is applied routinely[20,21]. 13 short linear stretches of polypeptide sequence, scattered throughout the viral proteome, were identified that exhibit exclusive reactivity towards ZIKV antisera

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Discussion

Conclusion