Development and characterization of serotype-specific monoclonal antibodies against the dengue virus-4 (DENV-4) non-stru
- PDF / 1,106,244 Bytes
- 12 Pages / 595.276 x 790.866 pts Page_size
- 59 Downloads / 204 Views
METHODOLOGY
Open Access
Development and characterization of serotype-specific monoclonal antibodies against the dengue virus-4 (DENV-4) non-structural protein (NS1) Tesfaye Gelanew and Elizabeth Hunsperger*
Abstract Background: Dengue, caused by one of the four serologically distinct dengue viruses (DENV-1 to − 4), is a mosquito-borne disease of serious global health significance. Reliable and cost-effective diagnostic tests, along with effective vaccines and vector-control strategies, are highly required to reduce dengue morbidity and mortality. Evaluation studies revealed that many commercially available NS1 antigen (Ag) tests have limited sensitivity to DENV-4 serotype compared to the other three serotypes. These studies indicated the need for development of new NS1 Ag detection test with improved sensitivity to DENV-4. An NS1 capture enzyme linked immunoassay (ELISA) specific to DENV-4 may improve the detection of DENV-4 cases worldwide. In addition, a serotype-specific NS1 Ag test identifies both DENV and the infecting serotype. Methods: In this study, we used a small-ubiquitin-like modifier (SUMO*) cloning vector to express a SUMO*-DENV-4 rNS1 fusion protein to develop NS1 DENV-4 specific monoclonal antibodies (MAbs). These newly developed MAbs were then optimized for use in an anti-NS1 DENV-4 capture ELISA. The serotype specificity and sensitivity of this ELISA was evaluated using (i) supernatants from DENV (1–4)-infected Vero cell cultures, (ii) rNS1s from all the four DENV (1–4) and, (iii) rNS1s of related flaviviruses (yellow fever virus; YFV and West Nile virus; WNV). Results: From the evaluation studies of the newly developed MAbs, we identified three DENV-4 specific anti-NS1 MAbs: 3H7A9, 8A6F2 and 6D4B10. Two of these MAbs were optimal for use in a DENV-4 serotype-specific NS1 capture ELISA: MAb 8A6F2 as the capture antibody and 6D4B10 as a detection antibody. Conclusion: This ELISA was sensitive and specific to DENV-4 with no cross-reactivity to other three DENV (1–3) serotypes and other heterologous flaviviruses. Taken together these data indicated that our MAbs are useful reagents for the development of DENV-4 immunodiagnostic tests. Keywords: Dengue virus 4, Non-structural protein 1, NS1, Diagnostic, flavivirus, Monoclonal antibody
Background Dengue is a serious disease of public importance with increasing worldwide spread. It is caused by an infection with any one of the four antigenically distinct dengue virus serotypes (DENV1–4). Current diagnosis of an acute DENV infection primarily relies on reverse transcription polymerase chain reaction (RT-PCR), a highly * Correspondence: [email protected] Dengue Branch, Division of Vector-Borne Diseases, National Center for Enteric and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention (CDC), San Juan, PR, USA
sophisticated test. To improve dengue case detection in dengue endemic countries, a highly sensitive, specific and simple rapid assay is needed to provide early support for patients and to accurately differentiate dengue from ot
Data Loading...
