Development and characterization of serotype-specific monoclonal antibodies against the dengue virus-4 (DENV-4) non-structural protein (NS1)

Abstract

BackgroundDengue, 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.MethodsIn 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).ResultsFrom 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.ConclusionThis 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.Electronic supplementary materialThe online version of this article (doi: 10.1186/s12985-018-0925-7) contains supplementary material, which is available to authorized users.

MCP-1 LEVELS AND ATYPICAL LYMPHOCYTES IN EARLY FEVER OF DENGUE VIRUS INFECTION WITH NON-STRUCTURAL PROTEIN 1 (NS-1) ANTIGEN TEST IN dr DARSONO HOSPITAL, PACITAN

“…This ELISA was sensitive and specific to DENV-4 with no cross-reactivity to other three DENV (1-3) serotypes and other heterologous flaviviruses. 10 The secondary infection with a different serotype occurs, the immune response can lead to the presentation of dengue fever is more severe in some cases. 32 The incubation period ranges from 3 to 14 days and symptoms usually develop between 4 and 7 days after vector bites.…”

“…Rapid tests such as the NS-1 enzyme-linked immunosorbent assay (ELISA) are commercially available for DENV with relatively good sensitivity and specificity. 10 Clinical diagnosis of dengue can be challenging, depending largely on what stage in the infection process a patient presents. Depending on the geographic region of the world, there can be a number of disease-causing pathogens or disease states that can mimic the disease spectrum arising from dengue infection.…”

“…NS-1 Rapid detection tests or enzyme-linked immunosorbent assay (ELISA) is available commercially for DENV was sensitive and specific to DENV-4 with no cross-reactivity to other three DENV (1-3) serotypes and other heterologous flaviviruses. 10 Ordering a dengue NS-1 antigen assay within the first week of symptom onset is one of the initial investigations recommended by the Ministry of Health, Singapore, for dengue infections. 35 NS1 test detects at the same time as viral RNA and before an antibody response, so the time to do the best examination is day 0 to day 4 and can be detected before the decrease in platelets.…”

“…DPO up to 18 at a concentration of NS-1 up to 50 μg/mL. 10 The NS-1 is a glycoprotein with two glycosylation sites that are conserved among flaviviruses. It is synthesized in the ER as a hydrophilic monomer but exists as a more hydrophobic homodimer.…”

Quantitative and Selective Surface Plasmon Resonance Response Based on a Reduced Graphene Oxide–Polyamidoamine Nanocomposite for Detection of Dengue Virus E-Proteins

“…There are four distinct serotypes of dengue viruses, i.e., DENV 1 to DENV 4, and each serotype consists 3 structural proteins (capsid, membrane, envelope (E) protein) and 7 non-structural proteins [41][42][43][44][45][46][47]. Currently, the detection of the NS1 antigen from the dengue virus is the basis for early diagnosis and an ideal diagnostic marker of disease progression when antibody levels are not detectable [48,49]. The setback of NS1 detection, however, is its low sensitivity in the diagnosis of secondary dengue infection (DENV 2) compared to primary dengue infection (DENV 1) [50][51][52]; therefore, we did not focus on DENV 2 detection.…”