Serving the Society Since 1986
Advanced Search
Volume 24 Issue 2
April 2009
    Citation: Min QING, Zhi-ming YUAN, Pei-Yong Shi. A Virus-type Specific Serological Diagnosis of Flavivirus Infection Using Virus-like Particles .VIROLOGICA SINICA, 2009, 24(2) : 136-145.  http://dx.doi.org/10.1007/s12250-009-3023-6
    shu

    A Virus-type Specific Serological Diagnosis of Flavivirus Infection Using Virus-like Particles

    • 1.

      Wadsworth Center, New York State Department of Health, 120 New Scotland Avenue, Albany, New York 12208, USA

    • 2.

      Stake Key Laboratory of Virology, Wuhan Institute of Virology Chinese Academy of Sciences, Wuhan 430071, China

    • 3.

      Department of Biomedical Sciences, University at Albany, State University of New York, Albany, New York, Albany, New York 12208, USA

    • Corresponding author: Pei-Yong Shi, pei_yong.shi@novartis.com
    • Received Date: 08 January 2009
      Accepted Date: 14 January 2009
      Available online: 01 April 2009
    • Many flaviviruses are emerging and reemerging pathogens, such as West Nile virus (WNV), dengue virus (DENV), yellow fever virus (YFV), and Japanese encephalitis virus. Serological assay is the dominant method for diagnosis of flavivirus infections in human. Because antibodies generated during flavivirus infections cross-react with other flavivirus members, plaque reduction neutralization test (PRNT) is the only available assay to determine the infecting flavivirus type. Since PRNT requires culturing raw viruses, it must be performed in biosafety level-3 or level-4 containment for many flaviviruses, and takes more than ten days to complete. To overcome these problems, we have developed flavivirus viral-like particles (VLPs) that could be used to replace raw viruses in the neutralization assay. The VLPs were prepared by trans packaging a luciferase-reporting replicon with viral structural proteins. This novel assay involves three simple steps: (i) VLPs from a panel of flaviviruses are incubated with flavivirus-infected sera at 37℃ for 1 h; (ii)the neutralized VLPs are used to infect Vero cells; and (iii) the infected cells are measured for luciferase activities at 22 h post-infection. The virus type whose VLP is most efficiently neutralized by the serum specimen (as quantified by the luciferase activities) is the etiologic agent. As a proof-of-concept, we show that a WNV-infected mouse serum neutralized the WNV VLP more efficiently and selectively than the DENV and YFV VLPs. Our results demonstrate that the VLP neutralization assay maintains the “gold standard” of the classic PRNT; importantly, it shortens the assay time from >10 days to <1 day, and can be performed in biosafety level-2 facility
    • 加载中

      1. Andrewes C H, Elford W J. 1933. Observations on anti-phage sera. I. 'The percentage law'. Br J Exp Pathol, 14: 367-376.

      2. Ansarah-Sobrinho C, Nelson S, Jost C A, et al.2008. Temperature-dependent production of pseudoinfectious dengue reporter virus particles by complementation. Virology, 381 (1): 67-74.
          doi: 10.1016/j.virol.2008.08.021

      3. Brinton M A. 2002. The molecular biology of West Nile Virus: a new invader of the western hemisphere. Annu Rev Microbiol, 56: 371-402.
          doi: 10.1146/annurev.micro.56.012302.160654

      4. Burke D S, Monath T P. 2001. Flaviviruses. In: Fields Virology (Knipe D M, Howley P M, Griffin D E, et al. eds), Philadelphia, PA: Lippincott, Williams, & Wilkins, p1043-1126.

      5. Chambers T J, Hahn C S, Galler R, et al. 1990. Flavivirus genome organization, expression, and replication. Annu Rev Microbiol, 44:649-688.
          doi: 10.1146/annurev.mi.44.100190.003245

      6. Colombage G, Hall R, Pavy, M, et al. 1998. DNA-based and alphavirus-vectored immunization with prM and E proteins elicits long-lived and protective immunity against the flavivirus, Murray Valley encephalitis virus. Virology, 250 (1): 151-163.
          doi: 10.1006/viro.1998.9357

      7. Deas T S, Binduga-Gajewska I, Tilgner M, et al. 2005. Inhibition of flavivirus infections by antisense oligomers specifically suppressing viral translation and RNA replication. J Virol, 79 (8):4599-4609.
          doi: 10.1128/JVI.79.8.4599-4609.2005

      8. Falconar A K. 1999. Identification of an epitope on the dengue virus membrane (M) protein defined by cross-protective monoclonal antibodies:Design of an improved epitope sequence based on common determinants present in both envelope (E and M) proteins. Arch Virol, 144 (12):2313-2330.
          doi: 10.1007/s007050050646

      9. Gubler D, Kuno G, Markoff L. 2007. Flaviviruses. In: Fields virology (Knipe D M, Howley P M. ed. ), 5th ed. , Philadelphia, PA: Lippincott, Williams, & Wilkins, . vol. 1, p1153-1253.

      10. Johnson A J, Martin D A, Karabatsos N, et al. 2000. Detection of anti-arboviral immunoglobulin G by using a monoclonal antibody-based capture enzyme-linked immunosorbent assay. J Clin Microbiol, 38 (5): 1827-1831.

      11. Jones C, Patkar C, Kuhn R. 2005. Construction and applications of yellow fever virus replicons. Virology, 331 (2): 247-259.
          doi: 10.1016/j.virol.2004.10.034

      12. Klasse P J, Sattentau Q J. 2001. Mechanisms of virus neutralization by antibody. Curr Top Microbiol Immunol, 260: 87-108.

      13. Kummerer B M, Rice C M. 2002. Mutations in the yellow fever virus nonstructural protein NS2A selectively block production of infectious particles. J Virol, 76 (10): 4773-4784.
          doi: 10.1128/JVI.76.10.4773-4784.2002

      14. Lindenbach B D, Rice C M. 2007. Flaviviridae: the virus 17 and their replication. In: Fields virology (Knipe D M, Howley P M, . Griffin D E, et al. ed. ), 4th ed. , Philadelphia, PA: Lippincott, Williams, & Wilkins, p991-1042.

      15. Liu W J, Chen H B, Khromykh A A. 2003. Molecular and functional analyses of Kunjin virus infectious cDNA clones demonstrate the essential roles for NS2A in virus assembly and for a nonconservative residue in NS3 in RNA replication. J Virol, 77 (14): 7804-7813.
          doi: 10.1128/JVI.77.14.7804-7813.2003

      16. Liu W, Wang X J, Mokhonov V V, et al. 2005. Inhibition of interferon signaling by the New York 99 strain and Kunjin subtype of West Nile virus involves blockage of STAT1 and STAT2 activation by nonstructural proteins. J Virol, 79 (3): 1934-1942.
          doi: 10.1128/JVI.79.3.1934-1942.2005

      17. Martin D A, Biggerstaff B J, Allen B, et al. 2002. Use of Immunoglobulin M Cross-Reactions in Differential Diagnosis of Human Flaviviral Encephalitis Infections in the United States. Clin Diag Lab Immunol, 9 (3): 544-549.

      18. Martin D A, Muth D A, Brown T, et al. 2000. Standardization of immunoglobulin M capture enzyme-linked immunosorbent assays for routine diagnosis of arboviral infections. J Clin Microbiol, 38 (5): 1823-1826.

      19. Mehlhop E, Ansarah-Sobrinho C, Johnson S, et al. 2007. Complement protein C1q inhibits antibody-dependent enhancement of flavivirus infection in an IgG subclassspecific manner. Cell Host Microbe, 2(6): 417-426.
          doi: 10.1016/j.chom.2007.09.015

      20. Munoz-Jordan J L, Sanchez-Burgos G G, Laurent-Rolle M, et al. 2003. Inhibition of interferon signaling by dengue virus. Proc Natl Acad Sci USA, 100 (24): 14333-14338.
          doi: 10.1073/pnas.2335168100

      21. Pierson T C, Sanchez M D, Puffer B A, et al. 2006. A rapid and quantitative assay for measuring antibody-mediated 18 neutralization of West Nile virus infection. Virology, 346 (1): 53-65.
          doi: 10.1016/j.virol.2005.10.030

      22. Pincus S, Mason P W, Konishi E, et al. 1992. Recombinant vaccinia virus producing the prM and E proteins of yellow fever virus protects mice from lethal yellow fever encephalitis. Virology, 187 (1): 290-297.
          doi: 10.1016/0042-6822(92)90317-I

      23. Puig-Basagoiti F, Tilgner M, Forshey B M, et al. 2006. Triaryl Pyrazoline Compound Inhibits Flavivirus RNA Replication. Antimicrob Agents Chemother, 50 (4): 1320-1329.
          doi: 10.1128/AAC.50.4.1320-1329.2006

      24. Puig-Basagoiti F, Deas T S, Ren P, et al. 2005. High-throughput assays using luciferase-expressing replicon, virus-like particle, and full-length virus for West Nile virus drug discovery. Antimicrob Agents Chemother, 49 (12): 4980-4988.
          doi: 10.1128/AAC.49.12.4980-4988.2005

      25. Shi P Y, Tilgner M, Lo M K. 2002. Construction and characterization of subgenomic replicons of New York strain of West Nile virus. Virology, 296(2): 219-233.
          doi: 10.1006/viro.2002.1453

      26. Tardei G, Ruta S, Chitu V, et al. 2000. Evaluation of immunoglobulin M (IgM) and IgG enzyme immunoassays in serologic diagnosis of West Nile virus infections. J Clin Microbiol, 38 (6):2232-2239.

      27. Vazquez S, Guzman M G, Guillen G, et al. 2002. Immune response to synthetic peptides of dengue prM protein. Vaccine, 20 (13-14):1823-1830.
          doi: 10.1016/S0264-410X(01)00515-1

    • 加载中

    Figures(4) / Tables(1)

    PDF

    Article Metrics

    Article views(4576) PDF downloads(16) Cited by()

    Related
    Proportional views

      A Virus-type Specific Serological Diagnosis of Flavivirus Infection Using Virus-like Particles

        Corresponding author: Pei-Yong Shi, pei_yong.shi@novartis.com
      • 1. Wadsworth Center, New York State Department of Health, 120 New Scotland Avenue, Albany, New York 12208, USA
      • 2. Stake Key Laboratory of Virology, Wuhan Institute of Virology Chinese Academy of Sciences, Wuhan 430071, China
      • 3. Department of Biomedical Sciences, University at Albany, State University of New York, Albany, New York, Albany, New York 12208, USA
      • Received Date: 08 January 2009
      • Accepted Date: 14 January 2009

      Abstract: Many flaviviruses are emerging and reemerging pathogens, such as West Nile virus (WNV), dengue virus (DENV), yellow fever virus (YFV), and Japanese encephalitis virus. Serological assay is the dominant method for diagnosis of flavivirus infections in human. Because antibodies generated during flavivirus infections cross-react with other flavivirus members, plaque reduction neutralization test (PRNT) is the only available assay to determine the infecting flavivirus type. Since PRNT requires culturing raw viruses, it must be performed in biosafety level-3 or level-4 containment for many flaviviruses, and takes more than ten days to complete. To overcome these problems, we have developed flavivirus viral-like particles (VLPs) that could be used to replace raw viruses in the neutralization assay. The VLPs were prepared by trans packaging a luciferase-reporting replicon with viral structural proteins. This novel assay involves three simple steps: (i) VLPs from a panel of flaviviruses are incubated with flavivirus-infected sera at 37℃ for 1 h; (ii)the neutralized VLPs are used to infect Vero cells; and (iii) the infected cells are measured for luciferase activities at 22 h post-infection. The virus type whose VLP is most efficiently neutralized by the serum specimen (as quantified by the luciferase activities) is the etiologic agent. As a proof-of-concept, we show that a WNV-infected mouse serum neutralized the WNV VLP more efficiently and selectively than the DENV and YFV VLPs. Our results demonstrate that the VLP neutralization assay maintains the “gold standard” of the classic PRNT; importantly, it shortens the assay time from >10 days to <1 day, and can be performed in biosafety level-2 facility

        HTML