A novel indirect ELISA for diagnosis of dengue fever

Abstract

Background & objectives:Dengue fever (DF) is associated with significant morbidity and mortality in the tropical and sub-tropical regions of the world. Since there are no effective antiviral drugs for treatment, clinicians often rely on the accurate diagnosis of dengue fever to begin supportive therapy at early stages of the illness. The objective of this study was to develop an in-house dengue virus serotype 2 (DENV-2) non-structural protein- 5 (NS5) based indirect ELISA.Methods:DENV-2 was raised in Vero cells and the viral proteins were separated and subsequently the NS5 protein was eluted. Serum samples from primary and secondary dengue fever patients; and acute and convalescent samples from Japanese encephalitis (JE) and West Nile virus (WNV) cases were used to validate the ELISA.Results:The assay was found to be 100 per cent specific in detecting DENV-2 specific antibodies from patient's serum. However, in terms of sensitivity, the assay could detect IgM antibodies only from 90 per cent of the primary dengue samples. The IgM/IgG ratio of the primary and secondary samples was 7.24 and 0.64, respectively.Interpretation & conclusions:The results indicate that the DENV-2 NS5 ELISA is dengue group specific and can be used to differentiate dengue infection from other circulating Flavivirus infections. This NS5 ELISA can also be used to distinguish between primary and secondary dengue fever on the basis of IgM/IgG ratios. Further studies with larger sample sizes and different DENV serotypes are required to validate the ELISA.

Molecular characterization of recombinant premembrane protein of dengue virus serotype‐2 for development of diagnostic assay

“…Aedes mosquito is the primary cause of transmission of dengue, Chikungunya, Yellow fever, and Zika virusrelated disease complications. Pakistan has been facing many outbreaks of dengue infection over the last three decades, with a high [12] number of deaths. Dengue infection is Pakistan's most prevalent vector-borne viral infection [13].…”

Linear and Continuous Flavivirus Epitopes From Naturally Infected Humans

“…In this review, we evaluated data related to 153 linear immunoreactive peptides from viruses within the Flavivirus genus, including the DENV, ZIKV, JEV, WNV, and TBEV, as reported in 29 manuscripts published from 1986 to 2020. Unsurprisingly, the largest number of peptides in the literature was defined for the DENV, especially for the DENV serotype 2 (Narayan et al, 2016), followed by the ZIKV (Vogt et al, 2011), DENV3 (Dong et al, 2012), DENV1 (Filgueira and Lannes, 2019), JEV (Petersen et al, 2013), WNV (Pierson and Diamond, 2018), and TBEV (Pierson and Diamond, 2018). Consequently, the reduced number of peptides described for other flaviviruses may be partly due to their lesser impact on human health and the existence of effective vaccines or methods of prophylaxis (i.e., for the JEV, TBEV, WNV, and YFV).…”

“…The antigenic potential of the nonstructural proteins is still not fully understood; thus, serological identification of antibodies directed against immunogenic peptides from these proteins may be intriguing. However, a number of studies have identified immunoreactive antibodies against these proteins, including elevated seroprevalence of anti-Ns4A during secondary infection, with Ns5 used as a specific antigen for serological detection (Se-Thoe et al, 1999;Wong et al, 2003;Narayan et al, 2016). Antibodies against Ns5 and other nonstructural proteins were observed during DENV infection, but their functional relevance is still unclear (Churdboonchart et al, 1991;Valdes et al, 2000).…”

Identification of B-Cell Epitopes with Potential to Serologicaly Discrimnate Dengue from Zika Infections

“…However, several studies involving epitopes of different non-structural DENV proteins either for diagnostic or vaccine purposes have been described. As an example, Narayan and co-workers [40] described an ELISA using the DENV2-derived NS5 protein as an antigen for serotype-specific serological detection. In recent studies aimed at proving which epitopes of DENV and ZIKV proteomes were responsible for the most effective responses in B and TCD8 + activation, researchers have reported NS3, NS4, and NS5 sequences as important response generators and, thus, possible vaccine or diagnostic targets—for both DENV and ZIKV [41].…”