An antigen capture enzyme-linked immunosorbent assay reveals high levels of the dengue virus protein NS1 in the sera of infected patients

Young, Paul R., Hilditch, Paige A., Bletchly, Cheryl and Halloran, Wendy (2000) An antigen capture enzyme-linked immunosorbent assay reveals high levels of the dengue virus protein NS1 in the sera of infected patients. Journal of Clinical Microbiology, 38 3: 1053-1057.


Author Young, Paul R.
Hilditch, Paige A.
Bletchly, Cheryl
Halloran, Wendy
Title An antigen capture enzyme-linked immunosorbent assay reveals high levels of the dengue virus protein NS1 in the sera of infected patients
Journal name Journal of Clinical Microbiology   Check publisher's open access policy
Publication date 2000-03
Sub-type Article
Volume number 38
Issue number 3
ISSN 0095-1137
Start page 1053
End page 1057
Total pages 5
Place of publication Washington, DC
Publisher American Society for Microbiology
Collection year 2000
Language eng
Subject C1
270303 Virology
730101 Infectious diseases
Abstract We describe the development of a capture enzyme-linked immunosorbent assay for the detection of the dengue virus nonstructural protein NS1. The assay employs rabbit polyclonal and monoclonal antibodies as the capture and detection antibodies, respectively. Immunoaffinity-purified NS1 derived from dengue 2 virus-infected cells was used as a standard to establish a detection sensitivity of approximately 4 ng/ml for an assay employing monoclonal antibodies recognizing a dengue 2 serotype-specific epitope. A number of serotype cross-reactive monoclonal antibodies were also shown to be suitable probes for the detection of NS1 expressed by the remaining three dengue virus serotypes. Examination of clinical samples demonstrated that the assay was able to detect NS1 with minimal interference from serum components at the test dilutions routinely used, suggesting that it could form the basis of a useful additional diagnostic test for dengue virus infection. Furthermore, quantitation of NS1 levels in patient sera may prove to be a valuable surrogate marker for viremia. Surprisingly high levels of NS1, as much as 15 mu g/ml, were found in acute-phase sera taken hom some of the patients experiencing serologically confirmed dengue 2 virus secondary infections but was not detected in the convalescent sera of these patients. In contrast, NS1 could not be detected in either acute-phase or convalescent serum samples taken from patients with serologically confirmed primary infection. The presence of high levels of secreted NS1 in the sera of patients experiencing secondary dengue virus infections, and in the context of an anamnestic antibody response, suggests that NS1 may contribute significantly to the formation of the circulating immune complexes that are suspected to play an important role in the pathogenesis of severe dengue disease.
Keyword Microbiology
Nonstructural Glycoprotein Ns1
Yellow-fever Virus
Monoclonal-antibodies
Molecular-biology
Immune-complexes
Rna Replication
Purification
Complement
Challenge
Epitopes
Q-Index Code C1

Document type: Journal Article
Sub-type: Article
Collections: School of Chemistry and Molecular Biosciences
Institute for Molecular Bioscience - Publications
 
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Created: Sun, 19 Aug 2007, 15:28:58 EST