Development and clinical evaluation of a highly accurate dengue NS1 rapid test: from the preparation of a soluble NS1 antigen to the construction of an RDT

Highlight
• DENV NS1 antigen is useful for the early confirmation of DENV infection.
• A soluble DENV NS1 protein is prepared with high antigenicity and purity.
• Monoclonal antibodies are generated against the DENV NS1 protein with high affinity.
• A highly accurate dengue NS1 rapid test is developed using anti-DENV NS1 mAbs.
• Clinical evaluation shows that the test detects 4 dengue serotypes with improved diagnostic performance.
• The test could offer a reliable point-of-care testing tool for early dengue detection.

Early diagnosis of dengue virus (DENV) is important. There are numerous products on the market claiming to detect DENV NS1, but these are not always reliable. In this study, a highly sensitive and accurate rapid diagnostic test (RDT) was developed using anti-dengue NS1 monoclonal antibodies. A recombinant NS1 protein was produced with high antigenicity and purity. Monoclonal antibodies were raised against this purified NS1 antigen. The RDT was constructed using a capturing (4A6A10, Kd = 7.512 ± 0.419 × 10−9) and a conjugating antibody (3E12E6, Kd = 7.032 ± 0.322 × 10−9). The diagnostic performance was evaluated with NS1-positive clinical samples collected from various dengue endemic countries and compared to SD BioLine Dengue NS1 Ag kit. The constructed RDT exhibited higher sensitivity (92.9%) with more obvious diagnostic performance than the commercial kit (83.3%). The specificity of constructed RDT was 100%. The constructed RDT could offer a reliable point-of-care testing tool for the early detection of dengue infections in remote areas and contribute to the control of dengue-related diseases.