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Clinical and vaccine immunology : CVI2007; 14(8); 1024-1031; doi: 10.1128/CVI.00051-07

Epitope-blocking enzyme-linked immunosorbent assay to differentiate west nile virus from Japanese encephalitis virus infections in equine sera.

Abstract: West Nile virus (WNV) is now widely distributed worldwide, except in most areas of Asia where Japanese encephalitis virus (JEV) is distributed. Considering the movement and migration of reservoir birds, there is concern that WNV may be introduced in Asian countries. Although manuals and guidelines for serological tests have been created in Japan in preparedness for the introduction of WNV, differential diagnosis between WNV and JEV may be complicated by antigenic cross-reactivities between these flaviviruses. Here, we generated a monoclonal antibody specific for the nonstructural protein 1 (NS1) of WNV and established an epitope-blocking enzyme-linked immunosorbent assay that can differentiate WNV from JEV infections in horse sera. Under conditions well suited for our assay system, samples collected from 95 horses in Japan (regarded as negative for WNV antibodies), including those collected from horses naturally infected with JEV, showed a mean inhibition value of 8.2% and a standard deviation (SD) of 6.5%. However, inhibition values obtained with serum used as a positive control (obtained after 28 days from a horse experimentally infected with WNV) in nine separate experiments showed a mean of 54.4% and an SD of 7.1%. We tentatively determined 27.6% (mean + 3 x SD obtained with 95 negative samples) as the cutoff value to differentiate positive from negative samples. Under this criterion, two horses experimentally infected with WNV were diagnosed as positive at 12 and 14 days, respectively, after infection.
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Publication Date: 2007-06-27 PubMed ID: 17596430PubMed Central: PMC2044481DOI: 10.1128/CVI.00051-07Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This study presents a new way to differentiate between West Nile virus and Japanese encephalitis virus infections in horses by using an epitope-blocking enzyme-linked immunosorbent assay.

Objective and Background of the Research

  • The objective of this research was to create a more specific diagnostic tool for distinguishing between West Nile virus (WNV) and Japanese encephalitis virus (JEV) in horse sera.
  • WNV is a viral infection that is commonly found across the globe while JEV is found mainly in Asia.
  • Considering the movement patterns of reservoir birds, there is a risk of WNV being introduced to Asian countries. This has raised concerns due to the potential for misdiagnosis owing to antigenic cross-reactivities between these two flaviviruses.

Method: Epitope-Blocking Enzyme-Linked Immunosorbent Assay

  • The researchers developed a monoclonal antibody specific to the nonstructural protein 1 (NS1) of WNV.
  • This was used to establish an epitope-blocking enzyme-linked immunosorbent (ELISA) assay.
  • The ELISA assay is a common diagnostic tool that measures the concentration of an analyte (a substance that is tested in the lab) in a liquid sample by using antibodies and color change.
  • In this context, the assay was used to differentiate between WNV infections and JEV infections in horse sera.

Results

  • Different conditions suitable for the assay system were tested with samples collected from 95 horses in Japan that were considered negative for WNV antibodies. Some of these samples were from horses naturally infected with JEV. The mean inhibition value obtained from these tests was 8.2%, with a standard deviation (SD) of 6.5%.
  • The researchers used serum as a positive control, taken from a horse that had been experimentally infected with WNV. The mean inhibition value from nine separate experiments with this serum was 54.4%, and the SD was 7.1%.
  • The cutoff value used to differentiate between positive and negative samples was tentatively set at 27.6%. This was calculated based on the mean inhibition value plus three times the standard deviation obtained through testing the negative samples.
  • When this criterion was applied, two horses that had been experimentally infected with WNV tested positive at 12 and 14 days after infection, respectively.

Conclusion

  • This study presents a specific and measurable method of differentiating WNV from JEV infections in horses.
  • The results suggest that the assay could be used for diagnosing West Nile virus in regions where Japanese encephalitis virus is endemic.

Cite This Article

APA
Kitai Y, Shoda M, Kondo T, Konishi E. (2007). Epitope-blocking enzyme-linked immunosorbent assay to differentiate west nile virus from Japanese encephalitis virus infections in equine sera. Clin Vaccine Immunol, 14(8), 1024-1031. https://doi.org/10.1128/CVI.00051-07

Publication

Clinical and vaccine immunology : CVI
ISSN: 1556-6811
NlmUniqueID: 101252125
Country: United States
Language: English
Volume: 14
Issue: 8
Pages: 1024-1031

Researcher Affiliations

Kitai, Yoko
  • Department of Health Sciences, Kobe University School of Medicine, 7-10-2 Tomogaoka, Suma-ku, Kobe 654-0142, Japan.
Shoda, Mizue
    Kondo, Takashi
      Konishi, Eiji

        MeSH Terms

        • Animals
        • Antibodies, Monoclonal / immunology
        • Antibodies, Viral / blood
        • Antibodies, Viral / immunology
        • Antibody Specificity
        • Chlorocebus aethiops
        • Encephalitis Virus, Japanese / classification
        • Encephalitis Virus, Japanese / immunology
        • Encephalitis, Japanese / diagnosis
        • Encephalitis, Japanese / veterinary
        • Encephalitis, Japanese / virology
        • Enzyme-Linked Immunosorbent Assay / methods
        • Epitopes / immunology
        • Horse Diseases / diagnosis
        • Horse Diseases / immunology
        • Horse Diseases / virology
        • Horses
        • Japanese Encephalitis Vaccines
        • Vero Cells
        • Viral Nonstructural Proteins / immunology
        • West Nile Fever / diagnosis
        • West Nile Fever / veterinary
        • West Nile Fever / virology
        • West Nile virus / classification
        • West Nile virus / immunology

        References

        This article includes 53 references
        1. Blitvich BJ, Fernandez-Salas I, Contreras-Cordero JF, Marlenee NL, Gonzalez-Rojas JI, Komar N, Gubler DJ, Calisher CH, Beaty BJ. Serologic evidence of West Nile virus infection in horses, Coahuila State, Mexico.. Emerg Infect Dis 2003 Jul;9(7):853-6.
          pmc: PMC3023419pubmed: 12890327doi: 10.3201/eid0907.030166google scholar: lookup
        2. Blitvich BJ, Marlenee NL, Hall RA, Calisher CH, Bowen RA, Roehrig JT, Komar N, Langevin SA, Beaty BJ. Epitope-blocking enzyme-linked immunosorbent assays for the detection of serum antibodies to west nile virus in multiple avian species.. J Clin Microbiol 2003 Mar;41(3):1041-7.
          pmc: PMC150274pubmed: 12624027doi: 10.1128/jcm.41.3.1041-1047.2003google scholar: lookup
        3. Blitvich BJ, Scanlon D, Shiell BJ, Mackenzie JS, Hall RA. Identification and analysis of truncated and elongated species of the flavivirus NS1 protein.. Virus Res 1999 Mar;60(1):67-79.
          pubmed: 10225275doi: 10.1016/s0168-1702(99)00003-9google scholar: lookup
        4. Burke DS, Monath TP. Flavivirus. p. 1043-1125. In D. M. Knipe and P. M. Howley (ed.), Fields virology, 4th ed. Lippincott Williams & Wilkins, Philadelphia, PA.
        5. Calisher CH, Gould EA. Taxonomy of the virus family Flaviviridae.. Adv Virus Res 2003;59:1-19.
          pubmed: 14696325doi: 10.1016/s0065-3527(03)59001-7google scholar: lookup
        6. Castillo-Olivares J, Wood J. West Nile virus infection of horses.. Vet Res 2004 Jul-Aug;35(4):467-83.
          pubmed: 15236677doi: 10.1051/vetres:2004022google scholar: lookup
        7. Dauphin G, Zientara S. West Nile virus: recent trends in diagnosis and vaccine development.. Vaccine 2007 Jul 26;25(30):5563-76.
          doi: 10.1016/j.vaccine.2006.12.005pubmed: 17292514google scholar: lookup
        8. Hall RA, Broom AK, Hartnett AC, Howard MJ, Mackenzie JS. Immunodominant epitopes on the NS1 protein of MVE and KUN viruses serve as targets for a blocking ELISA to detect virus-specific antibodies in sentinel animal serum.. J Virol Methods 1995 Feb;51(2-3):201-10.
          pubmed: 7738140doi: 10.1016/0166-0934(94)00105-pgoogle scholar: lookup
        9. Hall RA, Kay BH, Burgess GW, Clancy P, Fanning ID. Epitope analysis of the envelope and non-structural glycoproteins of Murray Valley encephalitis virus.. J Gen Virol 1990 Dec;71 ( Pt 12):2923-30.
          pubmed: 1703213doi: 10.1099/0022-1317-71-12-2923google scholar: lookup
        10. Hall RA, Scherret JH, Mackenzie JS. Kunjin virus: an Australian variant of West Nile?. Ann N Y Acad Sci 2001 Dec;951:153-60.
          pubmed: 11797773
        11. Hanna JN, Ritchie SA, Phillips DA, Shield J, Bailey MC, Mackenzie JS, Poidinger M, McCall BJ, Mills PJ. An outbreak of Japanese encephalitis in the Torres Strait, Australia, 1995.. Med J Aust 1996 Sep 2;165(5):256-60.
          pubmed: 8816682doi: 10.5694/j.1326-5377.1996.tb124960.xgoogle scholar: lookup
        12. Hayes EB, Gubler DJ. West Nile virus: epidemiology and clinical features of an emerging epidemic in the United States.. Annu Rev Med 2006;57:181-94.
          pubmed: 16409144doi: 10.1146/annurev.med.57.121304.131418google scholar: lookup
        13. Henchal EA, Gentry MK, McCown JM, Brandt WE. Dengue virus-specific and flavivirus group determinants identified with monoclonal antibodies by indirect immunofluorescence.. Am J Trop Med Hyg 1982 Jul;31(4):830-6.
          pubmed: 6285749doi: 10.4269/ajtmh.1982.31.830google scholar: lookup
        14. Ishikawa T, Konishi E. Mosquito cells infected with Japanese encephalitis virus release slowly-sedimenting hemagglutinin particles in association with intracellular formation of smooth membrane structures.. Microbiol Immunol 2006;50(3):211-23.
          pubmed: 16547419doi: 10.1111/j.1348-0421.2006.tb03788.xgoogle scholar: lookup
        15. Kobayashi M, Kurane I. Guidelines for the countermeasure to vector mosquitoes of West Nile Fever. National Institute of Infectious Diseases, Tokyo, Japan. (In Japanese.).
        16. Komar N. West Nile virus: epidemiology and ecology in North America.. Adv Virus Res 2003;61:185-234.
          pubmed: 14714433doi: 10.1016/s0065-3527(03)61005-5google scholar: lookup
        17. Komar O, Robbins MB, Klenk K, Blitvich BJ, Marlenee NL, Burkhalter KL, Gubler DJ, Gonzálvez G, Peña CJ, Peterson AT, Komar N. West Nile virus transmission in resident birds, Dominican Republic.. Emerg Infect Dis 2003 Oct;9(10):1299-302.
          pmc: PMC3033087pubmed: 14609467doi: 10.3201/eid0910.030222google scholar: lookup
        18. Konishi E. Monoclonal antibodies multireactive with parasite antigens produced by hybridomas generated from naive mice.. Parasitology 1997 Oct;115 ( Pt 4):387-93.
          pubmed: 9364565doi: 10.1017/s0031182097001509google scholar: lookup
        19. Konishi E, Fujii A. Dengue type 2 virus subviral extracellular particles produced by a stably transfected mammalian cell line and their evaluation for a subunit vaccine.. Vaccine 2002 Jan 15;20(7-8):1058-67.
          pubmed: 11803066doi: 10.1016/s0264-410x(01)00446-7google scholar: lookup
        20. Konishi E, Fujii A, Mason PW. Generation and characterization of a mammalian cell line continuously expressing Japanese encephalitis virus subviral particles.. J Virol 2001 Mar;75(5):2204-12.
          pmc: PMC114804pubmed: 11160724doi: 10.1128/jvi.75.5.2204-2212.2001google scholar: lookup
        21. Konishi E, Kosugi S, Imoto J. Dengue tetravalent DNA vaccine inducing neutralizing antibody and anamnestic responses to four serotypes in mice.. Vaccine 2006 Mar 15;24(12):2200-7.
          pubmed: 16316713doi: 10.1016/j.vaccine.2005.11.002google scholar: lookup
        22. Konishi E, Shoda M, Ajiro N, Kondo T. Development and evaluation of an enzyme-linked immunosorbent assay for quantifying antibodies to Japanese encephalitis virus nonstructural 1 protein to detect subclinical infections in vaccinated horses.. J Clin Microbiol 2004 Nov;42(11):5087-93.
          pmc: PMC525176pubmed: 15528700doi: 10.1128/jcm.42.11.5087-5093.2004google scholar: lookup
        23. Konishi E, Shoda M, Kondo T. Prevalence of antibody to Japanese encephalitis virus nonstructural 1 protein among racehorses in Japan: indication of natural infection and need for continuous vaccination.. Vaccine 2004 Mar 12;22(9-10):1097-103.
          pubmed: 15003636doi: 10.1016/j.vaccine.2003.10.001google scholar: lookup
        24. Konishi E, Shoda M, Kondo T. Analysis of yearly changes in levels of antibodies to Japanese encephalitis virus nonstructural 1 protein in racehorses in central Japan shows high levels of natural virus activity still exist.. Vaccine 2006 Jan 23;24(4):516-24.
          pubmed: 16129523doi: 10.1016/j.vaccine.2005.07.083google scholar: lookup
        25. Konishi E, Suzuki T. Ratios of subclinical to clinical Japanese encephalitis (JE) virus infections in vaccinated populations: evaluation of an inactivated JE vaccine by comparing the ratios with those in unvaccinated populations.. Vaccine 2002 Nov 22;21(1-2):98-107.
          pubmed: 12443667doi: 10.1016/s0264-410x(02)00433-4google scholar: lookup
        26. Koraka P, Zeller H, Niedrig M, Osterhaus AD, Groen J. Reactivity of serum samples from patients with a flavivirus infection measured by immunofluorescence assay and ELISA.. Microbes Infect 2002 Oct;4(12):1209-15.
          pubmed: 12467761doi: 10.1016/s1286-4579(02)01647-7google scholar: lookup
        27. Kozak M. Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes.. Cell 1986 Jan 31;44(2):283-92.
          pubmed: 3943125doi: 10.1016/0092-8674(86)90762-2google scholar: lookup
        28. Kuno G. Serodiagnosis of flaviviral infections and vaccinations in humans.. Adv Virus Res 2003;61:3-65.
          pubmed: 14714429doi: 10.1016/s0065-3527(03)61001-8google scholar: lookup
        29. Kurane I. West Nile fever. p. 104-105. In Japan Medical Association and Ministry of Health, Labour and Welfare (ed.). Guidelines for diagnosis and treatments of infectious diseases 2004. Igakushoin, Tokyo, Japan. (In Japanese.).
        30. Lai CJ, Monath TP. Chimeric flaviviruses: novel vaccines against dengue fever, tick-borne encephalitis, and Japanese encephalitis.. Adv Virus Res 2003;61:469-509.
          pubmed: 14714441doi: 10.1016/s0065-3527(03)61013-4google scholar: lookup
        31. Lian WC, Liau MY, Mao CL. Diagnosis and genetic analysis of Japanese encephalitis virus infected in horses.. J Vet Med B Infect Dis Vet Public Health 2002 Oct;49(8):361-5.
          pubmed: 12449242doi: 10.1046/j.1439-0450.2002.00509.xgoogle scholar: lookup
        32. Lin YL, Chen LK, Liao CL, Yeh CT, Ma SH, Chen JL, Huang YL, Chen SS, Chiang HY. DNA immunization with Japanese encephalitis virus nonstructural protein NS1 elicits protective immunity in mice.. J Virol 1998 Jan;72(1):191-200.
          pmc: PMC109364pubmed: 9420215doi: 10.1128/jvi.72.1.191-200.1998google scholar: lookup
        33. Lindenbach BD, Rice CM. Flaviviridae: the viruses and their replication. p. 991-1041. In D. M. Knipe and P. M. Howley (ed.), Fields virology, 4th ed. Lippincott Williams & Wilkins, Philadelphia, PA.
        34. Lvov DK, Butenko AM, Gromashevsky VL, Larichev VP, Gaidamovich SY, Vyshemirsky OI, Zhukov AN, Lazorenko VV, Salko VN, Kovtunov AI, Galimzyanov KM, Platonov AE, Morozova TN, Khutoretskaya NV, Shishkina EO, Skvortsova TM. Isolation of two strains of West Nile virus during an outbreak in southern Russia, 1999.. Emerg Infect Dis 2000 Jul-Aug;6(4):373-6.
          pmc: PMC2640908pubmed: 10905970doi: 10.3201/eid0604.000408google scholar: lookup
        35. Mackenzie JS, Gubler DJ, Petersen LR. Emerging flaviviruses: the spread and resurgence of Japanese encephalitis, West Nile and dengue viruses.. Nat Med 2004 Dec;10(12 Suppl):S98-109.
          pubmed: 15577938doi: 10.1038/nm1144google scholar: lookup
        36. McLean RG, Ubico SR, Bourne D, Komar N. West Nile virus in livestock and wildlife.. Curr Top Microbiol Immunol 2002;267:271-308.
          pubmed: 12082994doi: 10.1007/978-3-642-59403-8_14google scholar: lookup
        37. Ministry of Agriculture, Forestry and Fisheries and Ministry of Health, Labour and Welfares, Japan. Manuals for prevention from West Nile virus infections. Ministry of Agriculture, Forestry and Fisheries, Tokyo, Japan. (In Japanese.).
        38. Minton C. What have we learned from banding and flagging waders in Australia?. p. 116-142. In P. Straw (ed.), Status and conservation of shorebirds in the East Asian-Australasian flyway. Australasian Wader Studies Group and Wetlands International—Oceania, Sydney, Australia.
        39. Morales MA, Barrandeguy M, Fabbri C, Garcia JB, Vissani A, Trono K, Gutierrez G, Pigretti S, Menchaca H, Garrido N, Taylor N, Fernandez F, Levis S, Enría D. West Nile virus isolation from equines in Argentina, 2006.. Emerg Infect Dis 2006 Oct;12(10):1559-61.
          pmc: PMC3290965pubmed: 17176571doi: 10.3201/eid1210.060852google scholar: lookup
        40. Petersen LR, Marfin AA. Shifting epidemiology of Flaviviridae.. J Travel Med 2005 Apr;12 Suppl 1:S3-11.
          pubmed: 16225801doi: 10.2310/7060.2005.12052google scholar: lookup
        41. Platonov AE. West Nile encephalitis in Russia 1999-2001: were we ready? Are we ready?. Ann N Y Acad Sci 2001 Dec;951:102-16.
          pubmed: 11797768doi: 10.1111/j.1749-6632.2001.tb02689.xgoogle scholar: lookup
        42. ProMED-mail. Confirmation of circulation of West Nile virus in birds in the Far Eastern region of Russia. ProMED-mail 24 January 2005. Archive no. 20050124.0260. http://www.promedmail.org.
        43. ProMED-mail. West Nile virus in birds in the Vladivostok Region. ProMED-mail 30 June 2004. Archive no. 20040630.1744. http://www.promedmail.org.
        44. Putnak R, Porter K, Schmaljohn C. DNA vaccines for flaviviruses.. Adv Virus Res 2003;61:445-68.
          pubmed: 14714440doi: 10.1016/s0065-3527(03)61012-2google scholar: lookup
        45. Reed KD, Meece JK, Henkel JS, Shukla SK. Birds, migration and emerging zoonoses: west nile virus, lyme disease, influenza A and enteropathogens.. Clin Med Res 2003 Jan;1(1):5-12.
          pmc: PMC1069015pubmed: 15931279doi: 10.3121/cmr.1.1.5google scholar: lookup
        46. Sejvar JJ, Marfin AA. Manifestations of West Nile neuroinvasive disease.. Rev Med Virol 2006 Jul-Aug;16(4):209-24.
          pubmed: 16906589doi: 10.1002/rmv.501google scholar: lookup
        47. Shi PY, Wong SJ. Serologic diagnosis of West Nile virus infection.. Expert Rev Mol Diagn 2003 Nov;3(6):733-41.
          pubmed: 14628901doi: 10.1586/14737159.3.6.733google scholar: lookup
        48. Solomon T, Vaughn DW. Pathogenesis and clinical features of Japanese encephalitis and West Nile virus infections.. Curr Top Microbiol Immunol 2002;267:171-94.
          pubmed: 12082989doi: 10.1007/978-3-642-59403-8_9google scholar: lookup
        49. Turell MJ, Sardelis MR, O'Guinn ML, Dohm DJ. Potential vectors of West Nile virus in North America.. Curr Top Microbiol Immunol 2002;267:241-52.
          pubmed: 12082992doi: 10.1007/978-3-642-59403-8_12google scholar: lookup
        50. Usuku S, Noguchi Y, Takasaki T. Newly developed TaqMan assay to detect West Nile viruses in a wide range of viral strains.. Jpn J Infect Dis 2004 Jun;57(3):129-30.
          pubmed: 15218227
        51. Williams DT, Daniels PW, Lunt RA, Wang LF, Newberry KM, Mackenzie JS. Experimental infections of pigs with Japanese encephalitis virus and closely related Australian flaviviruses.. Am J Trop Med Hyg 2001 Oct;65(4):379-87.
          pubmed: 11693888doi: 10.4269/ajtmh.2001.65.379google scholar: lookup
        52. Wong SJ, Boyle RH, Demarest VL, Woodmansee AN, Kramer LD, Li H, Drebot M, Koski RA, Fikrig E, Martin DA, Shi PY. Immunoassay targeting nonstructural protein 5 to differentiate West Nile virus infection from dengue and St. Louis encephalitis virus infections and from flavivirus vaccination.. J Clin Microbiol 2003 Sep;41(9):4217-23.
          pmc: PMC193845pubmed: 12958248doi: 10.1128/jcm.41.9.4217-4223.2003google scholar: lookup
        53. Wu SJ, Grouard-Vogel G, Sun W, Mascola JR, Brachtel E, Putvatana R, Louder MK, Filgueira L, Marovich MA, Wong HK, Blauvelt A, Murphy GS, Robb ML, Innes BL, Birx DL, Hayes CG, Frankel SS. Human skin Langerhans cells are targets of dengue virus infection.. Nat Med 2000 Jul;6(7):816-20.
          pubmed: 10888933doi: 10.1038/77553google scholar: lookup

        Citations

        This article has been cited 20 times.
        1. Beck C, Lowenski S, Durand B, Bahuon C, Zientara S, Lecollinet S. Improved reliability of serological tools for the diagnosis of West Nile fever in horses within Europe. PLoS Negl Trop Dis 2017 Sep;11(9):e0005936.
          doi: 10.1371/journal.pntd.0005936pubmed: 28915240google scholar: lookup
        2. Ti J, Li Z, Li X, Lu Y, Diao Y, Li F. Identification of one B-cell epitope from NS1 protein of duck Tembusu virus with monoclonal antibodies. PLoS One 2017;12(7):e0181177.
          doi: 10.1371/journal.pone.0181177pubmed: 28746401google scholar: lookup
        3. Galula JU, Chang GJ, Chuang ST, Chao DY. Establishment of an Algorithm Using prM/E- and NS1-Specific IgM Antibody-Capture Enzyme-Linked Immunosorbent Assays in Diagnosis of Japanese Encephalitis Virus and West Nile Virus Infections in Humans. J Clin Microbiol 2016 Feb;54(2):412-22.
          doi: 10.1128/JCM.02469-15pubmed: 26659204google scholar: lookup
        4. Cleton NB, Godeke GJ, Reimerink J, Beersma MF, Doorn HR, Franco L, Goeijenbier M, Jimenez-Clavero MA, Johnson BW, Niedrig M, Papa A, Sambri V, Tami A, Velasco-Salas ZI, Koopmans MP, Reusken CB. Spot the difference-development of a syndrome based protein microarray for specific serological detection of multiple flavivirus infections in travelers. PLoS Negl Trop Dis 2015 Mar;9(3):e0003580.
          doi: 10.1371/journal.pntd.0003580pubmed: 25767876google scholar: lookup
        5. Beck C, Jimenez-Clavero MA, Leblond A, Durand B, Nowotny N, Leparc-Goffart I, Zientara S, Jourdain E, Lecollinet S. Flaviviruses in Europe: complex circulation patterns and their consequences for the diagnosis and control of West Nile disease. Int J Environ Res Public Health 2013 Nov 12;10(11):6049-83.
          doi: 10.3390/ijerph10116049pubmed: 24225644google scholar: lookup
        6. Yamanaka A, Kotaki T, Konishi E. A mouse monoclonal antibody against dengue virus type 1 Mochizuki strain targeting envelope protein domain II and displaying strongly neutralizing but not enhancing activity. J Virol 2013 Dec;87(23):12828-37.
          doi: 10.1128/JVI.01874-13pubmed: 24049185google scholar: lookup
        7. Hua RH, Liu LK, Chen ZS, Li YN, Bu ZG. Comprehensive Mapping Antigenic Epitopes of NS1 Protein of Japanese Encephalitis Virus with Monoclonal Antibodies. PLoS One 2013;8(6):e67553.
          doi: 10.1371/journal.pone.0067553pubmed: 23825668google scholar: lookup
        8. Hobson-Peters J. Approaches for the development of rapid serological assays for surveillance and diagnosis of infections caused by zoonotic flaviviruses of the Japanese encephalitis virus serocomplex. J Biomed Biotechnol 2012;2012:379738.
          doi: 10.1155/2012/379738pubmed: 22570528google scholar: lookup
        9. De Filette M, Ulbert S, Diamond M, Sanders NN. Recent progress in West Nile virus diagnosis and vaccination. Vet Res 2012 Mar 1;43(1):16.
          doi: 10.1186/1297-9716-43-16pubmed: 22380523google scholar: lookup
        10. Sun E, Zhao J, Liu N, Yang T, Xu Q, Qin Y, Bu Z, Yang Y, Lunt RA, Wang L, Wu D. Comprehensive mapping of common immunodominant epitopes in the West Nile virus nonstructural protein 1 recognized by avian antibody responses. PLoS One 2012;7(2):e31434.
          doi: 10.1371/journal.pone.0031434pubmed: 22347477google scholar: lookup
        11. Yeh JY, Lee JH, Park JY, Seo HJ, Moon JS, Cho IS, Kim HP, Yang YJ, Ahn KM, Kyung SG, Choi IS, Lee JB. A diagnostic algorithm to serologically differentiate West Nile virus from Japanese encephalitis virus infections and its validation in field surveillance of poultry and horses. Vector Borne Zoonotic Dis 2012 May;12(5):372-9.
          doi: 10.1089/vbz.2011.0709pubmed: 22217162google scholar: lookup
        12. Hirota J, Shimoji Y, Shimizu S. New sensitive competitive enzyme-linked immunosorbent assay using a monoclonal antibody against nonstructural protein 1 of West Nile virus NY99. Clin Vaccine Immunol 2012 Feb;19(2):277-83.
          doi: 10.1128/CVI.05382-11pubmed: 22190400google scholar: lookup
        13. Sun EC, Ma JN, Liu NH, Yang T, Zhao J, Geng HW, Wang LF, Qin YL, Bu ZG, Yang YH, Lunt RA, Wang LF, Wu DL. Identification of two linear B-cell epitopes from West Nile virus NS1 by screening a phage-displayed random peptide library. BMC Microbiol 2011 Jul 6;11:160.
          doi: 10.1186/1471-2180-11-160pubmed: 21729328google scholar: lookup
        14. Hua RH, Chen NS, Qin CF, Deng YQ, Ge JY, Wang XJ, Qiao ZJ, Chen WY, Wen ZY, Liu WX, Hu S, Bu ZG. Identification and characterization of a virus-specific continuous B-cell epitope on the PrM/M protein of Japanese Encephalitis Virus: potential application in the detection of antibodies to distinguish Japanese Encephalitis Virus infection from West Nile Virus and Dengue Virus infections. Virol J 2010 Sep 22;7:249.
          doi: 10.1186/1743-422X-7-249pubmed: 20858291google scholar: lookup
        15. Kitai Y, Kondo T, Konishi E. Complement-dependent cytotoxicity assay for differentiating West Nile virus from Japanese encephalitis virus infections in horses. Clin Vaccine Immunol 2010 May;17(5):875-8.
          doi: 10.1128/CVI.00217-09pubmed: 20237201google scholar: lookup
        16. Suzuki R, Fayzulin R, Frolov I, Mason PW. Identification of mutated cyclization sequences that permit efficient replication of West Nile virus genomes: use in safer propagation of a novel vaccine candidate. J Virol 2008 Jul;82(14):6942-51.
          doi: 10.1128/JVI.00662-08pubmed: 18480453google scholar: lookup
        17. Chiou SS, Crill WD, Chen LK, Chang GJ. Enzyme-linked immunosorbent assays using novel Japanese encephalitis virus antigen improve the accuracy of clinical diagnosis of flavivirus infections. Clin Vaccine Immunol 2008 May;15(5):825-35.
          doi: 10.1128/CVI.00004-08pubmed: 18337381google scholar: lookup
        18. Konishi E, Kitai Y, Kondo T. Utilization of complement-dependent cytotoxicity to measure low levels of antibodies: application to nonstructural protein 1 in a model of Japanese encephalitis virus. Clin Vaccine Immunol 2008 Jan;15(1):88-94.
          doi: 10.1128/CVI.00347-07pubmed: 18032598google scholar: lookup
        19. Gularte JS, Strottmann DM, Spilki FR, Zanluca C. Diagnostic Techniques for Dengue Virus. Curr Top Microbiol Immunol 2026;447:179-225.
          doi: 10.1007/82_2025_307pubmed: 40495084google scholar: lookup
        20. Ishikawa T, Narita K, Matsuyama K, Masuda M. Dissemination of the Flavivirus Subgenomic Replicon Genome and Viral Proteins by Extracellular Vesicles. Viruses 2024 Mar 28;16(4).
          doi: 10.3390/v16040524pubmed: 38675867google scholar: lookup
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