Abstract
Dengue virus infection is a common tropical disease which often occurs without being detected. These asymptomatic cases provide information in relation to the manifestation of immunological aspects. In this study, we developed an ELISA method to compare neutralizing effects of dengue prM and E antibodies between dengue patients and their asymptomatic household members. Recombinant D2 premembrane (prM) was constructed, cloned, and tested for antigenicity. The recombinant protein was purified and tested with controls by using an indirect ELISA method. Positive dengue serum samples with their asymptomatic pair were then carried out onto the developed ELISA. In addition, commercially available recombinant envelope (E) protein was used to develop an ELISA which was tested with the same set of serum samples in the prM ELISA. Asymptomatic individuals showed preexisting heterotypic neutralizing antibodies. The recombinant prM was antigenically reactive in the developed ELISA. Dengue patients had higher prM and E antibodies compared to their household members. Our study highlights the neutralizing antibodies levels with respect to dengue prM and E between dengue patients and asymptomatic individuals.
Highlights
Dengue virus (DENV) is a Flavivirus with four serotypes (DENV1–4)
Samples were considered as dengue positive if (1) DENV nucleic acid was detected; (2) DENV NS1 antigen was present; or (3) (i) DENV IgM seroconversion occurred in paired sera, (iii) dengue total antibodies had a fourfold rise in titers in paired sera, or (iv) a combination of the above
Our observation shows a larger occurrence of homologous DENV infections as observed by the higher percentage of Identification AH023 AH023 AH071 AH078 AH090 AH100 AH101 AH103 AH147 AH148 AH155 AH155 AH166 AH192 AH226 AH232 AH232 AH240 KH015 KH049 KH076 KH114 KH121 KH125 KH144 KH145 KH148 KH149 KH157 KH162 KH165 KH166 KH168 KH204 KH217
Summary
Dengue virus (DENV) is a Flavivirus with four serotypes (DENV1–4). Approximately, 3.6 billion people who are about 55% of the world’s population across the globe are at risk of being infected with dengue [1]. A recent annual report suggested that there are 390 million dengue infections that occur yearly of which about 96 million represent dengue fever (DF), dengue hemorrhagic fever (DHF), and dengue shock syndrome (DSS), whereas the other 300 million represent mild or asymptomatic cases [2]. Infection by a particular serotype confers a lifelong immunity against the homologous serotype but only limited cross-protection to the remaining three serotypes [3]. Primary DENV infections are often asymptomatic and will generate immunity to the homologous strain. The presence of cross-reactive, nonneutralizing antibodies generated during a primary infection has been suggested to enhance the pathogenicity of subsequent infections via the process of antibody-dependent enhancement (ADE) [5]. The occurrence of ADE could substantially increase the risk of manifesting severe dengue during subsequent infections especially in the asymptomatic
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