A nonstructural protein 1 capture enzyme-linked immunosorbent assay specific for dengue viruses

• Published in: PloS one Vol. 18; no. 5; p. e0285878
• Main Authors: Lim, Pei-Yin, Ramapraba, Appanna, Loy, Thomas, Rouers, Angeline, Thein, Tun-Linn, Leo, Yee-Sin, Burton, Dennis R., Fink, Katja, Wang, Cheng-I
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 18.05.2023; Public Library of Science (PLoS)
• Subjects: Analysis; Antibodies; Antibodies, Viral; Antigens; Assaying; B cells; Biological markers; Biology and life sciences; Care and treatment; Cloning; Control; Cross Reactions; Dengue; Dengue fever; Dengue Virus; Dengue viruses; Diagnosis; Engineering and Technology; Enzyme-Linked Immunosorbent Assay; Enzymes; Epitopes; Ethylenediaminetetraacetic acid; Health aspects; Human subjects; Humans; Identification and classification; Infection; Infections; Medical research; Medicine and Health Sciences; Medicine, Experimental; Methods; Physical Sciences; Plasma; Proteins; Research and Analysis Methods; Review boards; Sensitivity and Specificity; Serotypes; Testing; Vector-borne diseases; Viral antibodies; Viral Nonstructural Proteins; Viruses; West Nile virus; Zika Virus; Zika Virus Infection; Singapore
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0285878

Dengue non-structural protein (NS1) is an important diagnostic marker during the acute phase of infection. Because NS1 is partially conserved across the flaviviruses, a highly specific DENV NS-1 diagnostic test is needed to differentiate dengue infection from Zika virus (ZIKV) infection. In this study, we characterized three newly isolated antibodies against NS1 (A2, D6 and D8) from a dengue-infected patient and a previously published human anti-NS1 antibody (Den3). All four antibodies recognized multimeric forms of NS1 from different serotypes. A2 bound to NS1 from DENV-1, -2, and -3, D6 bound to NS1 from DENV-1, -2, and -4, and D8 and Den3 interacted with NS1 from all four dengue serotypes. Using a competition ELISA, we found that A2 and D6 bound to overlapping epitopes on NS1 whereas D8 recognized an epitope distinct from A2 and D6. In addition, we developed a capture ELISA that specifically detected NS1 from dengue viruses, but not ZIKV, using Den3 as the capture antibody and D8 as the detecting antibody. This assay detected NS1 from all the tested dengue virus strains and dengue-infected patients. In conclusion, we established a dengue-specific capture ELISA using human antibodies against NS1. This assay has the potential to be developed as a point-of-care diagnostic tool.