title: Dengue reporter virus particles for measuring neutralizing antibodies against each of the four dengue serotypes.
abstract: The lack of reliable, high-throughput tools for characterizing anti-dengue virus (DENV) antibodies in large numbers of serum samples has been an obstacle in understanding the impact of neutralizing antibodies on disease progression and vaccine efficacy. A reporter system using pseudoinfectious DENV reporter virus particles (RVPs) was previously developed by others to facilitate the genetic manipulation and biological characterization of DENV virions. In the current study, we demonstrate the diagnostic utility of DENV RVPs for measuring neutralizing antibodies in human serum samples against all four DENV serotypes, with attention to the suitability of DENV RVPs for large-scale, long-term studies. DENV RVPs used against human sera yielded serotype-specific responses and reproducible neutralization titers that were in statistical agreement with Plaque Reduction Neutralization Test (PRNT) results. DENV RVPs were also used to measure neutralization titers against the four DENV serotypes in a panel of human sera from a clinical study of dengue patients. The high-throughput capability, stability, rapidity, and reproducibility of assays using DENV RVPs offer advantages for detecting immune responses that can be applied to large-scale clinical studies of DENV infection and vaccination.
authors.affiliation: Integral Molecular, Inc., Philadelphia, Pennsylvania, United States of America.
authors.forename: Kimberly
authors.initials: K
authors.lastname: Mattia
authors.forename: Bridget A
authors.initials: BA
authors.lastname: Puffer
authors.forename: Katherine L
authors.initials: KL
authors.lastname: Williams
authors.forename: Ritela
authors.initials: R
authors.lastname: Gonzalez
authors.forename: Meredith
authors.initials: M
authors.lastname: Murray
authors.forename: Emily
authors.initials: E
authors.lastname: Sluzas
authors.forename: Dan
authors.initials: D
authors.lastname: Pagano
authors.forename: Sandya
authors.initials: S
authors.lastname: Ajith
authors.forename: Megan
authors.initials: M
authors.lastname: Bower
authors.forename: Eli
authors.initials: E
authors.lastname: Berdougo
authors.forename: Eva
authors.initials: E
authors.lastname: Harris
authors.forename: Benjamin J
authors.initials: BJ
authors.lastname: Doranz
mesh_terms: D057134:Antibodies, Neutralizing / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D002460:Cell Line; D003716:Dengue Virus / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D006801:Humans; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014771:Virion / Q000276:immunology* / Q000378:metabolism*
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral
references.citation: J Virol Methods. 2006 Jun;134(1-2):74-85
references.pmid: 16417930
references.citation: Am J Trop Med Hyg. 1982 May;31(3 Pt 1):548-55
references.pmid: 6177259
references.citation: Am J Trop Med Hyg. 2009 Jan;80(1):61-5
references.pmid: 19141841
references.citation: J Infect Dis. 2004 Mar 15;189(6):990-1000
references.pmid: 14999601
references.citation: Pediatr Infect Dis J. 2007 Sep;26(9):850-2
references.pmid: 17721386
references.citation: Virology. 2008 Nov 10;381(1):67-74
references.pmid: 18801552
references.citation: J Immunol. 1967 Aug;99(2):291-6
references.pmid: 4961907
references.citation: Virology. 2005 Jan 20;331(2):247-59
references.pmid: 15629769
references.citation: Virology. 2005 Mar 30;334(1):28-40
references.pmid: 15749120
references.citation: Clin Microbiol Rev. 1990 Oct;3(4):376-96
references.pmid: 2224837
references.citation: Virology. 2006 Mar 1;346(1):53-65
references.pmid: 16325883
references.citation: Nat Med. 2004 Dec;10(12 Suppl):S98-109
references.pmid: 15577938
references.citation: J Virol. 2004 Jan;78(1):531-8
references.pmid: 14671135
references.citation: Am J Trop Med Hyg. 2000 Jul-Aug;63(1-2):5-11
references.pmid: 11357995
references.citation: Am J Trop Med Hyg. 1983 Jan;32(1):154-6
references.pmid: 6824120
references.citation: Am J Trop Med Hyg. 2009 Aug;81(2):287-92
references.pmid: 19635885
references.citation: Nature. 1977 Feb 24;265(5596):739-41
references.pmid: 404559
references.citation: Proc Natl Acad Sci U S A. 2001 Feb 27;98(5):2670-5
references.pmid: 11226297
references.citation: J Exp Med. 2003 Apr 7;197(7):823-9
references.pmid: 12682107
references.citation: J Virol. 2006 Feb;80(3):1290-301
references.pmid: 16415006
references.citation: J Biomed Sci. 2008 Jan;15(1):15-27
references.pmid: 17768670
references.citation: PLoS Pathog. 2011 Jun;7(6):e1002111
references.pmid: 21738473
references.citation: Am J Trop Med Hyg. 2009 Nov;81(5):825-33
references.pmid: 19861618
references.citation: J Clin Invest. 2004 Apr;113(7):946-51
references.pmid: 15057297
references.citation: J Clin Microbiol. 2007 Nov;45(11):3777-80
references.pmid: 17804661
references.citation: Am J Trop Med Hyg. 1982 Jul;31(4):830-6
references.pmid: 6285749
references.citation: J Virol. 2006 Dec;80(23):11418-31
references.pmid: 16928749
references.citation: Proc Natl Acad Sci U S A. 2006 Sep 19;103(38):14234-9
references.pmid: 16966609
references.citation: J Virol. 2003 Aug;77(16):8924-33
references.pmid: 12885909
references.citation: Am J Trop Med Hyg. 2008 Jul;79(1):115-22
references.pmid: 18606774
references.citation: Science. 1988 Jan 29;239(4839):476-81
references.pmid: 3277268
references.citation: Am J Trop Med Hyg. 1989 Apr;40(4):444-51
references.pmid: 2712199