Irene Bosch1,2,3, Ankita Reddy1,2, Helena de Puig2,4, Juan E Ludert5, Federico Perdomo-Celis6, Carlos F Narváez6, Alice Versiani7, Diana Fandos2,8, Mauricio L Nogueira9, Mohit Singla10, Rakesh Lodha10, Guruprasad R Medigeshi11, Ivette Lorenzana12, Hugo Vicente Ralde13, Margarita Gélvez-Ramírez14, Luis A Villar14, Megan Hiley2, Laura Mendoza1, Nol Salcedo1, Bobby Brooke Herrera1,15, Lee Gehrke1,2,16. 1. E25Bio, Cambridge, Massachusetts, United States of America. 2. Institute for Medical Engineering & Science, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America. 3. Department of Medicine, Mount Sinai School of Medicine, New York, New York, United States of America. 4. Wyss Institute for Biologically Inspired Engineering, Harvard Medical School, Boston, Massachusetts, United States of America. 5. Departamento de Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV-IPN), Ciudad de México, México. 6. Programa de Medicina, Facultad de Salud, Universidad Surcolombiana, Neiva, Colombia. 7. Department of Infectious and Parasitic Diseases, Faculdade de Medicina de São José do Rio Preto, SP, Brazil. 8. Institut Químic de Sarrià, Universitat Ramon Llull, Barcelona, Spain. 9. Faculdade de Medicina de São José do Rio Preto (FAMERP), São José do Rio Preto, Brazil. 10. Department of Paediatrics, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, India. 11. Translational Health Science and Technology Institute, Faridabad, India. 12. Instituto de Investigación en Microbiología, Universidad Nacional Autónoma de Honduras, Tegucigalpa, Honduras. 13. Facultad de Medicina, Universidad Autónoma de Guadalajara, Guadalajara, Mexico. 14. Universidad Industrial de Santander and AEDES Network, Bucaramanga, Santander, Colombia. 15. Department of Immunology and Infectious Diseases, Harvard T.H. Chan School of Public Health, Boston, Massachusetts, United States of America. 16. Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts, United States of America.
Abstract
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.
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 DENVinfections, 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.
Authors: Thiago B Murari; Paulo Ferreira; Hugo Saba; Marcelo A Moret; Aloísio S Nascimento Filho Journal: Sci Rep Date: 2021-06-04 Impact factor: 4.379