Multicenter evaluation of a single-device dengue antibody/antigen rapid test for simultaneous detection of NS1, IgM,...

Dengue is a major global health burden in tropical and subtropical regions, where timely diagnosis is crucial for clinical management and outbreak control. Although rapid diagnostic tests (RDTs) are widely used at the point of care, their performance varies considerably. This study evaluated the diagnostic accuracy of the ASSURE Dengue Ab/Ag Rapid Test across three independent settings: Singapore, India, and Bangladesh, using reference enzyme-linked immunosorbent assays (ELISAs) and Conformité Européenne (CE)-marked RDTs. In Singapore, 430 characterized serum/plasma samples (141 positive and 289 negative) were tested against Panbio ELISAs and compared with the SD Standard Q Dengue Duo. In India, 300 prospective samples were tested against Panbio ELISAs. In Bangladesh, 85 retrospective samples were evaluated against commercial RDTs, ELISAs, and PCR. In Singapore, ASSURE achieved 100% sensitivity and specificity, outperforming SD Standard Q (sensitivity, 46.81%; specificity, 99.31%). In India, perfect concordance with Panbio ELISAs was observed across non-structural protein 1 (NS1), immunoglobulin M (IgM), and immunoglobulin G (IgG). In Bangladesh, NS1 detection was consistently strong (sensitivity, 100%; specificity, 88.14%-96.00%, depending on the comparator), while antibody detection showed lower sensitivity (IgM, 45.45%; IgG, 42.50%) but high specificity (91.07%-100%). Overall, the ASSURE Dengue Ab/Ag Rapid Test demonstrated superior diagnostic performance compared with CE-marked ELISAs and RDTs, particularly for NS1 antigen detection, and maintained accuracy across multicenter evaluations. These findings support its use as a reliable point-of-care tool for early dengue diagnosis.IMPORTANCEDengue fever is a mosquito-borne viral infection that affects millions annually. Early diagnosis is critical to reduce complications and improve patient outcomes. While rapid diagnostic tests (RDTs) are widely used, their reliability remains inconsistent. We evaluated the ASSURE Dengue Ab/Ag Rapid Test in Singapore, India, and Bangladesh, comparing it to established laboratory-based enzyme-linked immunosorbent assays and other commercial RDTs. Across studies, the test consistently achieved high accuracy, particularly for non-structural protein 1 antigen detection, which is crucial for identifying acute dengue infection. In contrast, the antibody components (immunoglobulin M and immunoglobulin G) showed variable sensitivity, highlighting the well-documented weaknesses and challenges that dengue tests currently face, particularly in antibody detection. Taken together, these results demonstrate that ASSURE is a dependable point-of-care test, especially for early-phase dengue diagnosis, and can improve patient management and outbreak response in endemic regions.