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Serotype-specific detection of dengue viruses in a nonstructural protein 1-based enzyme-linked immunosorbent assay validated with a multi-national cohort
- Source :
- PLoS Neglected Tropical Diseases, Vol 14, Iss 6, p e0008203 (2020), PLoS Neglected Tropical Diseases
- Publication Year :
- 2020
- Publisher :
- Public Library of Science (PLoS), 2020.
-
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.<br />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 (ELISA) 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 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.
- Subjects :
- RNA viruses
0301 basic medicine
Serotype
Physiology
viruses
RC955-962
Viral Nonstructural Proteins
Dengue virus
Pathology and Laboratory Medicine
Antibodies, Viral
medicine.disease_cause
Biochemistry
Geographical locations
Dengue fever
Cohort Studies
Dengue
0302 clinical medicine
Immune Physiology
Arctic medicine. Tropical medicine
Medicine and Health Sciences
Medicine
Enzyme-Linked Immunoassays
Immune System Proteins
biology
Antibodies, Monoclonal
virus diseases
Bioassays and Physiological Analysis
Infectious Diseases
Medical Microbiology
Viral Pathogens
Viruses
Cohort
Pathogens
Antibody
Public aspects of medicine
RA1-1270
Brazil
Research Article
Cohort study
Asia
medicine.drug_class
Immunology
030231 tropical medicine
India
Enzyme-Linked Immunosorbent Assay
Research and Analysis Methods
Serogroup
Monoclonal antibody
Microbiology
Sensitivity and Specificity
Antibodies
03 medical and health sciences
Immune system
Diagnostic Medicine
Animals
Humans
Immunoassays
Microbial Pathogens
Enzyme Assays
Flaviviruses
business.industry
Organisms
Public Health, Environmental and Occupational Health
Biology and Life Sciences
Proteins
Central America
Dengue Virus
South America
biochemical phenomena, metabolism, and nutrition
medicine.disease
Virology
Mice, Inbred C57BL
Latin America
030104 developmental biology
Honduras
North America
Immunologic Techniques
biology.protein
People and places
Biochemical Analysis
business
Subjects
Details
- Language :
- English
- ISSN :
- 19352735 and 19352727
- Volume :
- 14
- Issue :
- 6
- Database :
- OpenAIRE
- Journal :
- PLoS Neglected Tropical Diseases
- Accession number :
- edsair.doi.dedup.....2a4a38f070f92562666b1f7caef6c87d