A commercial anti-dengue virus (anti-DENV) indirect IgG enzyme-linked immunosorbent assay (ELISA) for serological diagnosis was evaluated for its utility in determining previous DENV exposure in U.S. travelers. The Boston Area Travel Medicine Network clinics used Focus Diagnostics anti-DENV IgG ELISA to measure anti-DENV IgG antibodies in 591 pretravel specimens from U.S. residents who had traveled to countries where dengue is endemic. When using the manufacturer's index cutoff value for this ELISA, false-positive results were observed that overestimated the perceived past DENV exposure in U.S. travelers. Validation of 121 of these anti-DENV IgG results by plaque reduction neutralization test (PRNT) was used for receiver operating characteristic (ROC) curve optimization of the index cutoff value from 1 to 3.0, improving the specificity of the anti-DENV IgG ELISA from 24% to 95.7%. Additionally, previous vaccination with yellow fever virus contributed to 52.8% of the false-positive rate in the anti-DENV IgG ELISA results. Optimization of the cutoff value of the anti-DENV IgG ELISA provided better interpretation and confidence in the results and eliminated the need for confirmation by PRNT. The travel history of U.S. travelers was also useful for categorizing these travelers into groups for analysis of previous DENV exposure.
Dengue is a major, global health problem, with an estimated 2.5 billion people at risk of contracting the disease and over 50 million infections annually (1). Dengue virus (family Flaviviridae, genus Flavivirus) is mosquito-borne with four antigenically distinct serotypes (DENV-1 to -4). There is sufficient antigenic homology among the four DENV serotypes and within the genus Flavivirus to create cross-reactivity in many immunoassays. This cross-reactivity is a diagnostic challenge when testing samples in regions where multiple DENV serotypes circulate and other flaviviruses cocirculate. Hence, immunoassays, such as the IgM enzyme-linked immunosorbent assay (ELISA) and the IgG ELISA, often require testing using a second method, such as a plaque reduction neutralization test (PRNT), to confirm the specificity of the antibody response (2).Anti-DENV IgG ELISAs are used to measure prevalence of previous infection by DENV and for conducting vaccine studies. The Boston Area Travel Medicine Network (BATMN) clinics used commercial anti-DENV IgG ELISAs to determine exposure to DENV in U.S. travelers; dengue is the leading cause of acute febrile illness in U.S. travelers returning from countries where DENV is endemic, as observed by GeoSentinal clinics (3). Among these patients, the Focus Diagnostics anti-DENV IgG ELISA had low specificity (24%) and high sensitivity (100%), as determined by the PRNT confirmatory test. Further evaluation of the cutoff value of the anti-DENV IgG ELISA was conducted in an attempt to increase the specificity of the test.
MATERIALS AND METHODSStudy. Travelers were enrolled from five travel clinics from the BATMN group in Boston, MA, from August 2008 through June 2009. Appro...