Volume 97, Issue 4
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645



Dengue virus (DENV) circulates in tropical and subtropical areas around the world, where it causes high morbidity and mortality. There is no effective treatment of infection, with supportive care being the only option. Furthermore, early detection and diagnosis are important to facilitate clinical decisions. In this study, seven monoclonal antibodies (mAbs) recognizing nonstructural protein 1 (NS1) of DENV were generated by hybridoma techniques. These antibodies can be divided into two groups: serotype-specific (DB6-1, DB12-3, and DB38-1) and nonspecific (consisting of antibodies DB16-1, DB20-6, DB29-1, and DB41-2). The B-cell epitopes of DB20-6 and DB29-1 were identified by phage display and site-directed mutagenesis, and its binding motif, WXXWGK, was revealed to correspond to amino acid residues 115–120 of the DENV-2 NS1 protein. A diagnostic platform, consisting of a serotype-specific capture antibody and a complex detection antibody, exhibited a detection limit of about 1 ng/mL, which is sufficient to detect NS1 in clinical serum samples from dengue patients. This diagnostic platform displayed better specificity and sensitivity than two examined commercial NS1 diagnostic platforms. In summary, our results indicate that these newly generated mAbs are suitable for detection of NS1 protein of DENV-2 in clinical samples.


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  1. Back AT, Lundkvist A, , 2013. Dengue viruses: an overview. Infect Ecol Epidemiol 3: 10.3402/iee.v3i0.19839. [Google Scholar]
  2. Li PC, Liao MY, Cheng PC, Liang JJ, Liu IJ, Chiu CY, Lin YL, Chang GJ, Wu HC, , 2012. Development of a humanized antibody with high therapeutic potential against dengue virus type 2. PLoS Negl Trop Dis 6: e1636.[Crossref] [Google Scholar]
  3. Malavige GN, Ogg GS, , 2013. T cell responses in dengue viral infections. J Clin Virol 58: 605611.[Crossref] [Google Scholar]
  4. Chuang YC, Lin YS, Liu CC, Liu HS, Liao SH, Shi MD, Lei HY, Yeh TM, , 2013. Factors contributing to the disturbance of coagulation and fibrinolysis in dengue virus infection. J Formos Med Assoc 112: 1217.[Crossref] [Google Scholar]
  5. Tang KF, Ooi EE, , 2012. Diagnosis of dengue: an update. Expert Rev Anti Infect Ther 10: 895907.[Crossref] [Google Scholar]
  6. Prestwood TR, May MM, Plummer EM, Morar MM, Yauch LE, Shresta S, , 2012. Trafficking and replication patterns reveal splenic macrophages as major targets of dengue virus in mice. J Virol 86: 1213812147.[Crossref] [Google Scholar]
  7. Prestwood TR, Prigozhin DM, Sharar KL, Zellweger RM, Shresta S, , 2008. A mouse-passaged dengue virus strain with reduced affinity for heparan sulfate causes severe disease in mice by establishing increased systemic viral loads. J Virol 82: 84118421.[Crossref] [Google Scholar]
  8. Jupatanakul N, Sim S, Dimopoulos G, , 2014. Aedes aegypti ML and Niemann-Pick type C family members are agonists of dengue virus infection. Dev Comp Immunol 43: 19.[Crossref] [Google Scholar]
  9. Zhou Y, Austin SK, Fremont DH, Yount BL, Huynh JP, de Silva AM, Baric RS, Messer WB, , 2013. The mechanism of differential neutralization of dengue serotype 3 strains by monoclonal antibody 8A1. Virology 439: 5764.[Crossref] [Google Scholar]
  10. Hu D, Di B, Ding X, Wang Y, Chen Y, Pan Y, Wen K, Wang M, Che X, , 2011. Kinetics of non-structural protein 1, IgM and IgG antibodies in dengue type 1 primary infection. Virol J 8: 47.[Crossref] [Google Scholar]
  11. Gutsche I, , et al., 2011. Secreted dengue virus nonstructural protein NS1 is an atypical barrel-shaped high-density lipoprotein. Proc Natl Acad Sci USA 108: 80038008.[Crossref] [Google Scholar]
  12. Young PR, Hilditch PA, Bletchly C, Halloran W, , 2000. An antigen capture enzyme-linked immunosorbent assay reveals high levels of the dengue virus protein NS1 in the sera of infected patients. J Clin Microbiol 38: 10531057. [Google Scholar]
  13. Winkler G, Maxwell SE, Ruemmler C, Stollar V, , 1989. Newly synthesized dengue-2 virus nonstructural protein NS1 is a soluble protein but becomes partially hydrophobic and membrane-associated after dimerization. Virology 171: 302305.[Crossref] [Google Scholar]
  14. Flamand M, Megret F, Mathieu M, Lepault J, Rey FA, Deubel V, , 1999. Dengue virus type 1 nonstructural glycoprotein NS1 is secreted from mammalian cells as a soluble hexamer in a glycosylation-dependent fashion. J Virol 73: 61046110. [Google Scholar]
  15. Hang VT, , et al., 2009. Diagnostic accuracy of NS1 ELISA and lateral flow rapid tests for dengue sensitivity, specificity and relationship to viraemia and antibody responses. PLoS Negl Trop Dis 3: e360.[Crossref] [Google Scholar]
  16. Sylvestre G, Gandini M, de Araujo JM, Kubelka CF, Lourenco-de-Oliveira R, Maciel-de-Freitas R, , 2014. Preliminary evaluation on the efficiency of the kit Platelia Dengue NS1 Ag-ELISA to detect dengue virus in dried Aedes aegypti: a potential tool to improve dengue surveillance. Parasit Vectors 7: 155.[Crossref] [Google Scholar]
  17. Jacobs MG, Robinson PJ, Bletchly C, Mackenzie JM, Young PR, , 2000. Dengue virus nonstructural protein 1 is expressed in a glycosyl-phosphatidylinositol-linked form that is capable of signal transduction. FASEB J 14: 16031610.[Crossref] [Google Scholar]
  18. Libraty DH, ., 2002. High circulating levels of the dengue virus nonstructural protein NS1 early in dengue illness correlate with the development of dengue hemorrhagic fever. J Infect Dis 186: 11651168.[Crossref] [Google Scholar]
  19. Bessoff K, Delorey M, Sun W, Hunsperger E, , 2008. Comparison of two commercially available dengue virus (DENV) NS1 capture enzyme-linked immunosorbent assays using a single clinical sample for diagnosis of acute DENV infection. Clin Vaccine Immunol 15: 15131518.[Crossref] [Google Scholar]
  20. Lapphra K, Sangcharaswichai A, Chokephaibulkit K, Tiengrim S, Piriyakarnsakul W, Chakorn T, Yoksan S, Wattanamongkolsil L, Thamlikitkul V, , 2008. Evaluation of an NS1 antigen detection for diagnosis of acute dengue infection in patients with acute febrile illness. Diagn Microbiol Infect Dis 60: 387391.[Crossref] [Google Scholar]
  21. De Decker S, Vray M, Sistek V, Labeau B, Enfissi A, Rousset D, Matheus S, , 2015. Evaluation of the diagnostic accuracy of a new dengue IgA capture assay (platelia dengue IgA capture, Bio-Rad) for dengue infection detection. PLoS Negl Trop Dis 9: e0003596.[Crossref] [Google Scholar]
  22. Koraka P, Burghoorn-Maas CP, Falconar A, Setiati TE, Djamiatun K, Groen J, Osterhaus AD, , 2003. Detection of immune-complex-dissociated nonstructural-1 antigen in patients with acute dengue virus infections. J Clin Microbiol 41: 41544159.[Crossref] [Google Scholar]
  23. Hermann LL, Thaisomboonsuk B, Poolpanichupatam Y, Jarman RG, Kalayanarooj S, Nisalak A, Yoon IK, Fernandez S, , 2014. Evaluation of a dengue NS1 antigen detection assay sensitivity and specificity for the diagnosis of acute dengue virus infection. PLoS Negl Trop Dis 8: e3193.[Crossref] [Google Scholar]
  24. Barniol J, , et al., 2011. Usefulness and applicability of the revised dengue case classification by disease: multi-centre study in 18 countries. BMC Infect Dis 11: 106.[Crossref] [Google Scholar]
  25. Castleberry JS, Mahon CR, , 2003. Dengue fever in the Western Hemisphere. Clin Lab Sci 16: 3438. [Google Scholar]
  26. Dussart P, Petit L, Labeau B, Bremand L, Leduc A, Moua D, Matheus S, Baril L, , 2008. Evaluation of two new commercial tests for the diagnosis of acute dengue virus infection using NS1 antigen detection in human serum. PLoS Negl Trop Dis 2: e280.[Crossref] [Google Scholar]
  27. Lima Mda R, Nogueira RM, Schatzmayr HG, dos Santos FB, , 2010. Comparison of three commercially available dengue NS1 antigen capture assays for acute diagnosis of dengue in Brazil. PLoS Negl Trop Dis 4: e738.[Crossref] [Google Scholar]
  28. Zhang Y, Bai J, Ying JY, , 2015. A stacking flow immunoassay for the detection of dengue-specific immunoglobulins in salivary fluid. Lab Chip 15: 14651471.[Crossref] [Google Scholar]
  29. Wang HK, Tsai CH, Chen KH, Tang CT, Leou JS, Li PC, Tang YL, Hsieh HJ, Wu HC, Cheng CM, , 2014. Cellulose-based diagnostic devices for diagnosing serotype-2 dengue fever in human serum. Adv Healthc Mater 3: 187196.[Crossref] [Google Scholar]
  30. Liu IJ, Chiu CY, Chen YC, Wu HC, , 2011. Molecular mimicry of human endothelial cell antigen by autoantibodies to nonstructural protein 1 of dengue virus. J Biol Chem 286: 97269736.[Crossref] [Google Scholar]
  31. Wu HC, Jung MY, Chiu CY, Chao TT, Lai SC, Jan JT, Shaio MF, , 2003. Identification of a dengue virus type 2 (DEN-2) serotype-specific B-cell epitope and detection of DEN-2-immunized animal serum samples using an epitope-based peptide antigen. J Gen Virol 84: 27712779.[Crossref] [Google Scholar]
  32. Alcon S, Talarmin A, Debruyne M, Falconar A, Deubel V, Flamand M, , 2002. Enzyme-linked immunosorbent assay specific to dengue virus type 1 nonstructural protein NS1 reveals circulation of the antigen in the blood during the acute phase of disease in patients experiencing primary or secondary infections. J Clin Microbiol 40: 376381.[Crossref] [Google Scholar]
  33. Avirutnan P, ., 2006. Vascular leakage in severe dengue virus infections: a potential role for the nonstructural viral protein NS1 and complement. J Infect Dis 193: 10781088.[Crossref] [Google Scholar]
  34. Srikiatkhachorn A, Kelley JF, , 2014. Endothelial cells in dengue hemorrhagic fever. Antiviral Res 109: 160170.[Crossref] [Google Scholar]
  35. Akey DL, ., 2014. Flavivirus NS1 structures reveal surfaces for associations with membranes and the immune system. Science 343: 881885.[Crossref] [Google Scholar]
  36. Schupbach J, Flepp M, Pontelli D, Tomasik Z, Luthy R, Boni J, , 1996. Heat-mediated immune complex dissociation and enzyme-linked immunosorbent assay signal amplification render p24 antigen detection in plasma as sensitive as HIV-1 RNA detection by polymerase chain reaction. AIDS 10: 10851090. [Google Scholar]
  37. Nadal D, Boni J, Kind C, Varnier OE, Steiner F, Tomasik Z, Schupbach J, , 1999. Prospective evaluation of amplification-boosted ELISA for heat-denatured p24 antigen for diagnosis and monitoring of pediatric human immunodeficiency virus type 1 infection. J Infect Dis 180: 10891095.[Crossref] [Google Scholar]
  38. Ledergerber B, Flepp M, Boni J, Tomasik Z, Cone RW, Luthy R, Schupbach J, , 2000. Human immunodeficiency virus type 1 p24 concentration measured by boosted ELISA of heat-denatured plasma correlates with decline in CD4 cells, progression to AIDS, and survival: comparison with viral RNA measurement. J Infect Dis 181: 12801288.[Crossref] [Google Scholar]
  39. Zhang B, Salieb-Beugelaar GB, Nigo MM, Weidmann M, Hunziker P, , 2015. Diagnosing dengue virus infection: rapid tests and the role of micro/nanotechnologies. Nanomedicine (Lond) 11: 17451761.[Crossref] [Google Scholar]

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  • Received : 02 Jan 2017
  • Accepted : 26 May 2017
  • Published online : 24 Jul 2017

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