Background: Extraordinary genetic diversity is a hallmark of dengue virus (DENV), with different genotypes within each of the four DENV serotypes capable of causing disease. The virus envelope (E) plays a critical role in the virus life cycle and it is the main target of the neutralizing antibody (NAb) responses. The E protein has accumulated intra-serotype genetic diversity with heterogeneous distribution worldwide, therefore evaluation of cross-genotype immunity is essential to track vaccine coverage in different parts of the globe. Takeda's live attenuated tetravalent dengue vaccine (TAK-003) is comprised of structural proteins from each serotype in an attenuated dengue virus type 2 (DENV-2) genomic backbone. In clinical trials TAK-003 has been found to be safe and well-tolerated, and to elicit long-lasting NAb responses against all vaccine-matched DENV serotypes irrespective of dengue serostatus at baseline. Methods and materials: Our goal was to evaluate the ability of post-vaccination samples to neutralize a broader selection of virus genotypes circulating in dengue endemic areas. A panel of genetically diverse DENV strains representative of historical (1956–2006) genotypes isolated in Asia and Latin America, were tested in a microneutralization assay. Results: Neutralization was observed across all genotypes and serotypes tested, with no significant genotype-specific differences in either dengue-seropositive or -seronegative vaccine recipients. Contemporary DENV strains isolated in Asia were sequenced and the E protein amino acid sequences aligned with the respective E proteins of the TAK-003 vaccine strains for each serotype. Structural modeling was used to predict location of amino acid differences with the largest potential for impact on the neutralization profile of the virus genotypes. A panel of DENV-specific reporter virus particles (RVPs) carrying E genes from contemporary strains were designed and constructed. Neutralization assays using these DENV RVPs and post-vaccination serum samples from Phase III clinical trials of TAK-003 are currently underway. Conclusion: These tests will enable the prediction of the ability of TAK-003 to elicit antibodies capable of neutralizing contemporary DENV strains circulating in areas of high endemicity.