Monoclonal antibody-based capture ELISA in the diagnosis of previous dengue infection

Abstract

BackgroundDengue is an important mosquito-borne disease. There is currently only one licensed vaccine for dengue prevention. The vaccine provides higher efficacy in pre-vaccination dengue-seropositive persons but a higher risk of subsequent more severe dengue in dengue-seronegative persons. It is recommended that the dengue vaccine may be given in dengue-seropositive individuals or as mass vaccination without individual pre-vaccination screening in areas where the dengue seroprevalence is > 80% in children aged 9 years. We evaluated a dengue specific immunoglobulin G monoclonal antibody-based capture enzyme-linked immunosorbent assay (MAb-ELISA) in the diagnosis of previous dengue infection using serum samples from the cohort study in Ratchaburi Province, Thailand.MethodsThe MAb-ELISA was compared to 70% plaque reduction neutralization test (PRNT70) in 453 serum samples from children aged 3–11 years in Ratchaburi Province, Thailand.ResultsThe sensitivity and specificity of MAb-ELISA at the positive to negative (P/N) ratio cut-off level of > 3 were both 0.91 in the diagnosis of previous dengue infection, compared to PRNT70. The false positivity was mainly in Japanese encephalitis (JE) seropositive subjects.ConclusionsThis research provides evidence that MAb-ELISA is useful for dengue seroprevalence study and dengue pre-vaccination screening. JE seropositivity was the major cause of false positive result in the study population.

Dengue virus seroprevalence study in Bangphae district, Ratchaburi, Thailand: A cohort study in 2012-2015

“…However, it was labor- and resource-intensive and time-consuming. Its interpretation requires technical expertise and tends to be complicated, especially after the secondary infection [ 14 , 24 ]. Performing ELISA is much simpler and less resource-intensive than performing PRNT; however, most of ELISA platforms are in-house developed and in absence of standardized information.…”

“…Furthermore, despite the fact that we used an in-house assay with limited validation [ 24 ], we demonstrated a high level of accuracy of IgG ELISA to determine DENV serostatus in serological evaluation. We recognize that the IgG ELISA used in this study was an in-house assay and is not available to be used in a non-research commercial setting for mass screening for pre-vaccination status.…”

“…This antibody detection test was based on in-house indirect sandwich dengue IgG ELISA technique [ 24 ]. 96-well ELISA plates were filled with 100 μl/well of a diluted dengue monoclonal antibody 2H2 1:50 in 0.018 M carbonate buffer.…”

“…On a subset (n = 640) of paired samples which showed a substantial rising of IgG titers over intervals, PRNT was performed to measure the neutralizing antibodies against DENV-1-4, as described previously [ 24 , 25 ]. PRNT 50 titer was defined as the highest reciprocal serum dilution where the virus infectivity was reduced by 50% compared with control.…”

Development of Monoclonal Antibodies and Antigen-Capture ELISA for Human Parechovirus Type 3

“…Nevertheless, we obtained mAbs specific to HPeV3, which recognized sites 133–159 aa and the 275–280 aa site of VP0, which was not reported earlier. Moreover, we also developed an ELISA for detecting HPeV3 antigen using two of our newly generated HPeV3-specific mAbs (#8 and #39), as mAb-based ELISA is highly specific and sensitive towards viral antigen detection [ 35 ]. Our Octet assay suggested that both mAbs show a high binding affinity to the full-length HPeV3-VP0 recombinant protein.…”