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High performance dengue virus antigen-based serotyping-NS1-ELISA (plus): A simple alternative approach to identify dengue virus serotypes in acute dengue specimens
- Source :
- PLoS Neglected Tropical Diseases, Vol 15, Iss 2, p e0009065 (2021), PLoS Neglected Tropical Diseases
- Publication Year :
- 2021
- Publisher :
- Public Library of Science (PLoS), 2021.
-
Abstract
- Dengue hemorrhagic fever (DHF) is caused by infection with dengue virus (DENV). Four different serotypes (DENV1-4) co-circulate in dengue endemic areas. The viral RNA genome-based reverse-transcription PCR (RT-PCR) is the most widely used method to identify DENV serotypes in patient specimens. However, the non-structural protein 1 (NS1) antigen as a biomarker for DENV serotyping is an emerging alternative method. We modified the serotyping-NS1-enzyme linked immunosorbent assay (stNS1-ELISA) from the originally established assay which had limited sensitivity overall and poor specificity for the DENV2 serotype. Here, four biotinylated serotype-specific antibodies were applied, including an entirely new design for detection of DENV2. Prediction of the infecting serotype of retrospective acute-phase plasma from dengue patients revealed 100% concordance with the standard RT-PCR method for all four serotypes and 78% overall sensitivity (156/200). The sensitivity of DENV1 NS1 detection was greatly improved (from 62% to 90%) by the addition of a DENV1/DENV3 sub-complex antibody pair. Inclusive of five antibody pairs, the stNS1-ELISA (plus) method showed an overall increased sensitivity to 85.5% (171/200). With the same clinical specimens, a commercial NS1 rapid diagnostic test (NS1-RDT) showed 72% sensitivity (147/200), significantly lower than the stNS1-ELISA (plus) performance. In conclusion, the stNS1-ELISA (plus) is an improved method for prediction of DENV serotype and for overall sensitivity. It could be an alternative assay not only for early dengue diagnosis, but also for serotype identification especially in remote resource-limited dengue endemic areas.<br />Author summary Four serotypes of DENV co-circulate in dengue endemic areas. Secondary infection with a different DENV serotype is beleived to involve with severe dengue disease. Standard laboratory diagnosis to identify DENV serotypes in dengue patient specimens is performed by sophisticated genome-based RT-PCR method with serotype-specific oligoprimers. We have previously established an alternative protein-based NS1 assay for DENV serotyping namely, a serotyping-NS1-ELISA (stNS1-ELISA), with the use of serotype-specific monoclonal antibodies (Mabs) to NS1 protein. Due to its unsatisfactory performance, the stNS1-ELISA was modified in this study. The biotinylated serotype-specific detection Mabs were introduced to enhance the overall sensitivity. A new DENV2-specific antibody was applied to improve DENV serotype identification. Prediction of infecting serotype from NS1-positive samples by our modified assay was 100% concordant with the standard RT-PCR method for all four serotypes. The overall sensitivity was greatly improved by an additional DENV1/DENV3 sub-complex antibody. This modified assay is efficient not only for early dengue diagnosis, but also for serotype identification in epidemiological studies and disease surveillance.
- Subjects :
- Serotype
RNA viruses
Viral Diseases
Physiology
viruses
RC955-962
Artificial Gene Amplification and Extension
Dengue virus
Viral Nonstructural Proteins
medicine.disease_cause
Antibodies, Viral
Pathology and Laboratory Medicine
Polymerase Chain Reaction
Biochemistry
Dengue fever
Dengue Fever
Dengue
Binding Analysis
Medical Conditions
Immune Physiology
Arctic medicine. Tropical medicine
Medicine and Health Sciences
Enzyme-Linked Immunoassays
Antigens, Viral
Rapid diagnostic test
Immune System Proteins
Reverse Transcriptase Polymerase Chain Reaction
Antibodies, Monoclonal
virus diseases
Infectious Diseases
Medical Microbiology
Viral Pathogens
Viruses
Biomarker (medicine)
Antibody
Pathogens
Public aspects of medicine
RA1-1270
Research Article
Neglected Tropical Diseases
Immunology
Enzyme-Linked Immunosorbent Assay
Biology
Serogroup
Research and Analysis Methods
Sensitivity and Specificity
Microbiology
Antibodies
Antigen
Diagnostic Medicine
medicine
Humans
Serotyping
Immunoassays
Molecular Biology Techniques
Microbial Pathogens
Molecular Biology
Chemical Characterization
Retrospective Studies
Dengue virus Antigen
Biology and life sciences
Flaviviruses
Public Health, Environmental and Occupational Health
Organisms
Proteins
Reverse Transcriptase-Polymerase Chain Reaction
Dengue Virus
medicine.disease
Tropical Diseases
Virology
Immunoglobulin M
Immunoglobulin G
biology.protein
Immunologic Techniques
Subjects
Details
- Language :
- English
- ISSN :
- 19352735 and 19352727
- Volume :
- 15
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- PLoS Neglected Tropical Diseases
- Accession number :
- edsair.doi.dedup.....2cfbc7c4b01722f27dcee8ddf12bd193