Dengue Virus Serotyping Based on Envelope and Membrane and Nonstructural Protein NS1 Serotype-Specific Capture Immunoglobulin M Enzyme-Linked Immunosorbent Assays

Abstract

Envelope and membrane (E/M) and nonstructural protein NS1 serotype-specific capture Immunoglobulin M (IgM) enzyme-linked immunosorbent assays (ELISAs) were developed to differentiate four dengue virus serotypes. A total of 93 anti-dengue virus IgM-positive serum samples collected between days 5 and 45 of illness from 59 confirmed dengue patients were analyzed. The results showed that positive serotype specificity could be identified for 86.1 and 47.6% of serum samples tested for E/M-specific IgM antibodies versus 83.3 and 42.9% of serum samples tested for NS1-specific IgM antibodies from patients with primary and secondary dengue virus infections, respectively. Dual analyses with both E/M and NS1 serotype-specific capture IgM ELISAs showed that positive serotype specificity could be correctly identified for 98.6 and 61.9% of all of the primary and secondary serum samples tested, respectively. These findings suggested that E/M and NS1 serotypespecific capture IgM ELISAs have the potential to be of use in dengue virus serotyping.

Nonstructural Protein 1-Specific Immunoglobulin M and G Antibody Capture Enzyme-Linked Immunosorbent Assays in Diagnosis of Flaviviral Infections in Humans

“…Based on the increase in P/N ratio value, the concentrations of antiprM/E antibodies were presumably 3-to 12-fold higher than those of anti-NS1 antibodies in the human serum depending on the timing of serum collection (date not shown). Furthermore, previous studies suggest that NS1 ELISAs can be used not only to differentiate natural infections from JEV vaccinations but also to differentiate primary and secondary dengue virus infections, with a serotyping capability (33,34). In the current study, we demonstrate that NS1 antibodies, regardless of isotype, are highly cross-reactive within the DENV serocomplex ( Fig.…”

Altered Enzyme-Linked Immunosorbent Assay Immunoglobulin M (IgM)/IgG Optical Density Ratios Can Correctly Classify All Primary or Secondary Dengue Virus Infections 1 Day after the Onset of Symptoms, when All of the Viruses Can Be Isolated

“…While variations in patients' immune responses during DV infections have thoroughly been described (2,10,17,18,19,20,21,23), subgroup classifications of these patients' antibody responses have not been established. In these previous studies, DV E/M protein-specific IgG antibodies were shown to rise before, or at the same time as, IgM antibodies during acute secondary flavivirus infections, while the IgM titers were usually lower than those generated during acute primary flavivirus infections (2,10,11,19,23).…”

“…Subsequently, other workers classified acute primary and secondary flavivirus infections by applying different discriminatory values directly to the IgM/IgG OD ratios (i.e., without converting the OD values to units). In these studies, acute primary and secondary flavivirus infections were identified using DV E/M protein-specific discriminatory IgM/IgG OD ratios of either Ͼ1.4 and Ͻ1.4, respectively, using patients' sera at 1/20 dilutions (14), or Ն1.2 and Ͻ1.2, respectively, using patients' sera at 1/100 dilutions (18,19,20).…”

Current Advances in Dengue Diagnosis

“…Dual analyses by both the E/M and the NS1 serotype-specific capture IgM ELISAs showed that positive serotype specificity could be correctly identified in 98.6 and 61.9% of all serum samples from patients with primary and secondary dengue virus infections, respectively (86). It is emphasized that equal amounts of each of the four dengue virus antigens should be added to the microtiter wells in order to obtain reliable results.…”