Serotype-specific detection of dengue viruses in a nonstructural protein 1-based enzyme-linked immunosorbent assay validated with a multi-national cohort

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Bosch, Irene ^{[1, 2, 3]} ; Reddy, Ankita ^{[2, 3]} ; de Puig, Helena ^{[2, 4]} ; Ludert, Juan E. ^[5] ; Perdomo-Celis, Federico ^[6] ; Narvaez, Carlos F. ^[6] ; Versiani, Alice ^[7] ; Fandos, Diana ^{[8, 2]} ; Nogueira, Mauricio L. ^[9] ; Singla, Mohit ^[10] ; Lodha, Rakesh ^[10] ; Medigeshi, Guruprasad R. ^[11] ; Lorenzana, Ivette ^[12] ; Ralde, Hugo Vicente ^[13] ; Gelvez-Ramirez, Margarita ^{[14, 15]} ; Villar, Luis A. ^{[14, 15]} ; Hiley, Megan ^[2] ; Mendoza, Laura ^[3] ; Salcedo, Nol ^[3] ; Herrera, Bobby Brooke ^{[16, 3]} ; Gehrke, Lee ^{[17, 2, 3]}

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^[1] Mt Sinai Sch Med, Dept Med, New York, NY 10029 - USA

^[2] MIT, Inst Med Engn & Sci, 77 Massachusetts Ave, Cambridge, MA 02139 - USA

^[3] E25Bio, Cambridge, MA 02139 - USA

^[4] Harvard Med Sch, Wyss Inst Biol Inspired Engn, Boston, MA 02115 - USA

^[5] Inst Politecn Nacl CINVESTAV IPN, Ctr Invest & Estudios Avanzados, Dept Infect & Patogenesis Mol, Mexico City, DF - Mexico

^[6] Univ Surcolombiana, Fac Salud, Programa Med, Neiva - Colombia

^[7] Fac Med Sao Jose Rio Preto, Dept Infect & Parasit Dis, Sao Jose Do Rio Preto, SP - Brazil

^[8] Univ Ramon Llull, Inst Quim Sarria, Barcelona - Spain

^[9] Fac Med Sao Jose Do Rio Preto FAMERP, Sao Jose Do Rio Preto - Brazil

^[10] All India Inst Med Sci, Dept Paediat, New Delhi - India

^[11] Translat Hlth Sci & Technol Inst, Faridabad - India

^[12] Univ Nacl Autnoma Honduras, Inst Invest Microbiol, Tegucigalpa - Honduras

^[13] Univ Autonoma Guadalajara, Fac Med, Guadalajara - Mexico

^[14] Univ Ind Santander, Bucaramanga - Colombia

^[15] AEDES Network, Bucaramanga - Colombia

^[16] Harvard TH Chan Sch Publ Hlth, Dept Immunol & Infect Dis, Boston, MA - USA

^[17] Harvard Med Sch, Dept Microbiol & Immunobiol, Boston, MA 02115 - USA

• Tipo de documento: Artigo Científico
• Fonte: PLoS Neglected Tropical Diseases; v. 14, n. 6 JUN 2020.
• Citações Web of Science: 0

Resumo

Author summary Dengue virus (DENV) infection is an increasingly significant threat to global health, with a yearly estimate of 390 million infections and an expected increasing burden with the rise of climate change and globalization. DENV is caused by one of the four serotypes (DENV-1-4), each of which have been associated with different immune responses and clinical manifestations. We developed a method to detect DENV serotypes by targeting the nonstructural 1 (NS1) antigen through an enzyme-linked immunosorbent-based assay with high sensitivity and specificity. We demonstrate that our high throughput mouse-derived antibody screening method selected for optimal test performance. The antibodies were integrated into an ELISA assay that can distinguish between the four different dengue serotypes by serotype-specific pairing. In addition, we provide a dengue universal antibody combination that enables pan-virus detection independently of the serotype. We use the ELISA in three different countries and calculate overall and site-specific sensitivities and specificities. The assay performs optimally when levels of viremia are high during the first five days of fever. Background Dengue virus (DENV) infections pose one of the largest global barriers to human health. The four serotypes (DENV 1-4) present different symptoms and influence immune response to subsequent DENV infections, rendering surveillance, risk assessments, and disease control particularly challenging. Early diagnosis and appropriate clinical management is critical and can be achieved by detecting DENV nonstructural protein 1 (NS1) in serum during the acute phase. However, few NS1-based tests have been developed that are capable of differentiating DENV serotypes and none are currently commercially available. Methodology/Principle findings We developed an enzyme-linked immunosorbent assay (ELISA) to distinguish DENV-1-4 NS1 using serotype-specific pairs of monoclonal antibodies. A total of 1,046 antibodies were harvested from DENV-immunized mice and screened for antigen binding affinity. ELISA clinical performance was evaluated using 408 polymerase chain reaction-confirmed dengue samples obtained from patients in Brazil, Honduras, and India. The overall sensitivity of the test for pan-DENV was 79.66% (325/408), and the sensitivities for DENV-1-4 serotyping were 79.1% (38/48), 80.41% (78/97), 100% (45/45), and 79.6% (98/123), respectively. Specificity reached 94.07-100%. Significance Our study demonstrates a robust antibody screening strategy that enabled the development of a serotype NS1-based ELISA with maximized specific and sensitive antigen binding. This sensitive and specific assay also utilized the most expansive cohort to date, and of which about half are from Latin America, a geographic region severely underrepresented in previous similar studies. This ELISA test offers potential enhanced diagnostics during the acute phase of infection to help guide patient care and disease control. These results indicate that this ELISA is a promising aid in early DENV-1-4 diagnosis and surveillance in regions of endemicity in addition to offer convenient monitoring for future vaccine interventions. (AU)

• Processo FAPESP: 18/17647-0 - Desenvolvimento de nanodispositivos diagnósticos sorológicos multiplex baseados em peptideos para a identificação diferencial de infecções por Dengue e Zika
• Beneficiário: Alice Freitas Versiani
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 13/21719-3 - Estudo epidemiológico da dengue (sorotipos 1 a 4) em coorte prospectiva de São José do Rio Preto, São Paulo, Brasil, durante 2014 a 2018
• Beneficiário: Maurício Lacerda Nogueira
• Modalidade de apoio: Auxílio à Pesquisa - Temático