Enzyme-linked immunosorbent assay using recombinant envelope protein 2 antigen for diagnosis of Chikungunya virus

Author(s)

Fumagalli, Marcilio Jorge ^[1] ; de Souza, William Marciel ^[1] ; Alves Esposito, Danillo Lucas ^[1] ; Silva, Angelica ^[1] ; Romeiro, Marilia Farignoli ^[1] ; Martinez, Edson Zangiacomi ^[2] ; Lopes da Fonseca, Benedito Antonio ^[1] ; Moraes Figueiredo, Luiz Tadeu ^[1]

Affiliation

^[1] Univ Sao Paulo, Ribeirao Preto Med Sch, Virol Res Ctr, Ribeirao Preto, SP - Brazil

^[2] Univ Sao Paulo, Ribeirao Preto Med Sch, Social Med, Ribeirao Preto, SP - Brazil

• Document type: Journal article
• Source: VIROLOGY JOURNAL; v. 15, JUL 24 2018.
• Web of Science Citations: 6

Abstract

Background: Chikungunya (CHIKV) virus is an important mosquito-borne virus causing outbreaks of acute febrile illness with arthropathy. The detection of specific antibodies against CHIKV is used for diagnosis after the acute viremic phase of the disease. However, a major challenge for serologic diagnosis of CHIKV and other alphaviruses is the cross-reactivity of antibodies to common antigens among these viruses. In the present study, we have developed an enzyme-linked immunosorbend assay using a recombinant envelope protein 2 of CHIKV produced in Escherichia coli system, as a capture antigen. Results: High titers (1600 to 12,800) of anti-CHIKV antibodies were detected in human sera analyzed by the CHIKV assay, suggesting it may detect low levels of the antibodies presence. On the other side, cross-reactivity was not observed in mouse hyperimmune sera to Mayaro virus and other alphaviruses analyzed by the CHIKV immunosorbend assay, suggesting it is a CHIKV-specific test. Fifty-nine human serum samples of CHIKV infection suspected cases were tested for immunoglobulin G (IgG) and M (IgM) antibodies detection using the CHIKV immunosorbend assay. A total of 44% (26/59) of samples were positive for IgG to CHIKV, determining 89.66% sensitivity and 100% specificity when the assay is compared to a CHIKV-specific neutralization assay. In addition, 40.6% (24/59) of samples were positive for IgM, determining 92.48% sensitivity and 79.04% specificity by a Bayesian method in the absence of a gold standard. Moreover, CHIKV immunosorbend assay showed similar sensibilities to a commercial immunochromatography assay (Lumiquick, USA) for CHIKV IgG and IgM detection. Conclusion: In short, we have developed a rapid, simple, specific and sensitive CHIKV immunosorbend assay for IgG and IgM detection and our results showed potential applicability on the diagnosis of infections by this virus. (AU)

• FAPESP's process: 16/01414-1 - Production of recombinant viral antigenic peptides and development of methods for serological diagnosis of infections by Chikungunya, Mayaro and Zika viruses
• Grantee: Marcilio Jorge Fumagalli
• Support Opportunities: Scholarships in Brazil - Master
• FAPESP's process: 14/02438-6 - Studies with Bunyaviridae that produce human disease
• Grantee: Luiz Tadeu Moraes Figueiredo
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 12/24150-9 - Research of virus in wild rodents, mosquitoes and ticks
• Grantee: William Marciel de Souza
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 17/13981-0 - Characterization, genomics and diagnostic of viruses with importance for public health in Brazil by high throughput sequencing
• Grantee: William Marciel de Souza
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 14/20851-8 - Study about Alphavirus and Flavivirus infections in samples of blood bank
• Grantee: Marilia Farignoli Romeiro
• Support Opportunities: Scholarships in Brazil - Doctorate