Document Detail

The development of a novel serotyping-NS1-ELISA to identify serotypes of dengue virus.
MedLine Citation:
PMID:  21277249     Owner:  NLM     Status:  MEDLINE    
BACKGROUND: Dengue virus (DENV), which causes mosquito-borne disease dengue hemorrhagic fever (DHF), consists of four serotypes co-circulating in endemic areas. Currently, DENV serotypes can be identified by laborious virus isolation followed by immunofluorescent assay and sophisticated RT-PCR.
OBJECTIVE: To establish a new assay designated as "serotyping-NS1-ELISA" to detect the NS1 protein and to identify DENV serotypes simultaneously.
STUDY DESIGN: The monoclonal antibodies (Mabs) against NS1 of each DENV serotype were produced and characterized for their serotype-specificity. To develop serotyping-NS1-ELISA, the selected serotype-specific anti-NS1 Mabs were applied to detect the NS1 antigen, which was previously captured by a flavivirus cross-reactive anti-NS1 Mab. Serotyping accuracy of the developed assay was validated with NS1 from DENV-infected cell culture supernatants and from well-characterized clinical specimens.
RESULTS: Of 30 anti-NS1 Mabs, 1 serotype-specific anti-NS1 Mab to each DENV serotype was selected based on NS1 capture ELISA results for developing the serotyping-NS1-ELISA. Using DENV-infected cell culture supernatants for validation, the selected antibodies were shown to be capable of differentiating four DENV serotypes. When acute phase plasma from DENV-infected patients was used for validation, 65 out of 85 specimens (76.5% overall sensitivity) were positive to one of the four serotypes developed in our assay. Interestingly, identification of DENV serotypes by our serotyping-NS1-ELISA was 100% accurate for DENV1, 3 and 4 and 82.4% for DENV2 as compared with standard RT-PCR. Assay specificity was 100% (90/90).
CONCLUSIONS: The developed serotyping-NS1-ELISA provides an alternative for simultaneous detection of DENV NS1 and identification of its serotype in acute patients' specimens. The assay would be applicable for dengue diagnosis and epidemiological studies.
Chunya Puttikhunt; Tanapan Prommool; Nathaporn U-thainual; Prapapun Ong-ajchaowlerd; Kroong Yoosook; Chutithorn Tawilert; Thaneeya Duangchinda; Aroonroong Jairangsri; Nattaya Tangthawornchaikul; Prida Malasit; Watchara Kasinrerk
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2011-02-01
Journal Detail:
Title:  Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology     Volume:  50     ISSN:  1873-5967     ISO Abbreviation:  J. Clin. Virol.     Publication Date:  2011 Apr 
Date Detail:
Created Date:  2011-03-15     Completed Date:  2011-07-28     Revised Date:  2011-08-25    
Medline Journal Info:
Nlm Unique ID:  9815671     Medline TA:  J Clin Virol     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  314-9     Citation Subset:  IM    
Copyright Information:
Copyright © 2011 Elsevier B.V. All rights reserved.
Medical Biotechnology Research Unit, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, Bangkok, Thailand.
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MeSH Terms
Antibodies, Monoclonal / immunology
Cross Reactions / immunology
Dengue Hemorrhagic Fever / virology
Dengue Virus / classification*,  immunology,  isolation & purification
Enzyme-Linked Immunosorbent Assay / methods*
Mice, Inbred BALB C
Serotyping / methods*
Viral Nonstructural Proteins / analysis*,  immunology
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/NS1 protein, dengue-1 virus; 0/Viral Nonstructural Proteins
Comment In:
J Clin Virol. 2011 Aug;51(4):289   [PMID:  21680239 ]

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