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

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 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). 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.

Detection of chikungunya virus‐specific IgM on laser‐cut paper‐based device using pseudo‐particles as capture antigen

“…Arboviruses are mainly prevalent in developing countries of tropical and subtropical areas and represent a serious public health concern for these countries . Diagnostic assays, such as reverse‐transcriptase polymerase chain reaction (RT‐PCR), plaque reduction neutralization test (PRNT) or enzyme‐linked immunosorbent assay (ELISA) can provide sensitive results and help to optimize patient care . However, these assays are time‐consuming, rarely affordable, and require infrastructure and qualified operators, making them accessible to relatively few low‐income countries …”

“…The development of rapid and affordable diagnostics tests can be an interesting alternative to the use of traditional diagnostic tests. Lateral flow assays (LFAs) for arboviruses diagnostics have been developed and are to date commercially available, both for the detection of viral proteins and for the serological of type M Immunoglobulins (IgM) and/or type G Immunoglobulins (IgG) . However, although the use of these tests is in principle suitable for low‐income countries, some issues have been reported, notably in terms of reliability, performance, quality, and regulatory approval .…”

“…10 Diagnostic assays, such as reversetranscriptase polymerase chain reaction (RT-PCR), plaque reduction neutralization test (PRNT) or enzyme-linked immunosorbent assay (ELISA) can provide sensitive results and help to optimize patient care. [11][12][13][14][15] However, these assays are time-consuming, rarely affordable, and require infrastructure and qualified operators, making them accessible to relatively few low-income countries. 16 The development of rapid and affordable diagnostics tests can be an interesting alternative to the use of traditional diagnostic tests.…”

“…Lateral flow assays (LFAs) for arboviruses diagnostics have been developed and are to date commercially available, both for the detection of viral proteins and for the serological of type M Immunoglobulins (IgM) and/or type G Immunoglobulins (IgG). 15,17,18 However, although the use of these tests is in principle suitable for low-income countries, 19 some issues have been reported, notably in terms of reliability, performance, quality, and regulatory approval. [20][21][22][23][24] Moreover, even with a regulated market price, they remain expensive and therefore relatively inaccessible to a great majority of the population living in the poorest countries.…”

Peptide Arrays of Three Collections of Human Sera from Patients Infected with Mosquito-Borne Viruses

“…The sequence similarity among many of these flaviviruses at the protein level contributes to the observed cross-reactivity in serological assays, which is especially high in the E protein and also present in the NS1 protein [11]. Although reports showing antibodies against other viral proteins are detectable [12], the E and NS1 proteins are the primary targets of the humoral anti-flavivirus immune response in humans [13][14][15].…”