Diagnosis of Zika Virus Infection by Peptide Array and Enzyme-Linked Immunosorbent Assay
Nischay Mishra,
Adrian Caciula,
Adam Price,
Riddhi Thakkar,
James Ng,
Lokendra V. Chauhan,
Komal Jain,
Xiaoyu Che,
Diego A. Espinosa,
Magelda Montoya Cruz,
Angel Balmaseda,
Eric H. Sullivan,
Jigar J. Patel,
Richard G. Jarman,
Jennifer L. Rakeman,
Christina T. Egan,
Chantal B. E. M. Reusken,
Marion P. G. Koopmans,
Eva Harris,
Rafal Tokarz,
Thomas Briese,
W. Ian Lipkin
Affiliations
Nischay Mishra
Center for Infection and Immunity, Mailman School of Public Health, Columbia University, New York, New York, USA
Adrian Caciula
Center for Infection and Immunity, Mailman School of Public Health, Columbia University, New York, New York, USA
Adam Price
Center for Infection and Immunity, Mailman School of Public Health, Columbia University, New York, New York, USA
Riddhi Thakkar
Center for Infection and Immunity, Mailman School of Public Health, Columbia University, New York, New York, USA
James Ng
Center for Infection and Immunity, Mailman School of Public Health, Columbia University, New York, New York, USA
Lokendra V. Chauhan
Center for Infection and Immunity, Mailman School of Public Health, Columbia University, New York, New York, USA
Komal Jain
Center for Infection and Immunity, Mailman School of Public Health, Columbia University, New York, New York, USA
Xiaoyu Che
Center for Infection and Immunity, Mailman School of Public Health, Columbia University, New York, New York, USA
Diego A. Espinosa
Division of Infectious Diseases and Vaccinology, School of Public Health, University of California, Berkeley, California, USA
Magelda Montoya Cruz
Division of Infectious Diseases and Vaccinology, School of Public Health, University of California, Berkeley, California, USA
Angel Balmaseda
Laboratorio Nacional de Virología, Centro Nacional de Diagnóstico y Referencia, Ministry of Health, Managua, Nicaragua
Eric H. Sullivan
Roche Madison, Madison, Wisconsin, USA
Jigar J. Patel
Roche Madison, Madison, Wisconsin, USA
Richard G. Jarman
Viral Diseases Branch, Walter Reed Army Institute of Research, Silver Spring, Maryland, USA
Jennifer L. Rakeman
Public Health Laboratory, New York City Department of Health and Mental Hygiene, New York, New York, USA
Christina T. Egan
Biodefense Laboratory, Wadsworth Center, New York State Department of Health, Albany, New York, USA
Chantal B. E. M. Reusken
WHO Collaborating Centre for Arbovirus and Viral Haemorrhagic Fever Reference and Research, Department of Viroscience, Erasmus University Medical Centre, Rotterdam, The Netherlands
Marion P. G. Koopmans
WHO Collaborating Centre for Arbovirus and Viral Haemorrhagic Fever Reference and Research, Department of Viroscience, Erasmus University Medical Centre, Rotterdam, The Netherlands
Eva Harris
Division of Infectious Diseases and Vaccinology, School of Public Health, University of California, Berkeley, California, USA
Rafal Tokarz
Center for Infection and Immunity, Mailman School of Public Health, Columbia University, New York, New York, USA
Thomas Briese
Center for Infection and Immunity, Mailman School of Public Health, Columbia University, New York, New York, USA
W. Ian Lipkin
Center for Infection and Immunity, Mailman School of Public Health, Columbia University, New York, New York, USA
ABSTRACT Zika 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). IMPORTANCE The emergence of Zika virus (ZIKV) as a teratogen is a profound challenge to global public health. Molecular diagnosis of infection is straightforward during the 3-week period when patients are viremic. However, serological diagnosis thereafter of historical exposure has been confounded by cross-reactivity. 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. These assays may be useful in guiding clinical management of mothers at risk for potential exposure to ZIKV and enable insights into the epidemiology of ZIKV infections.
