A group-specific, indirect sandwich ELISA for the detection of equine encephalosis virus antigen

被引:14
作者
Crafford, JE
Guthrie, AJ
van Vuuren, M
Mertens, PPC
Burroughs, JN
Howell, PG
Hamblin, C
机构
[1] Univ Pretoria, Fac Vet Sci, Dept Vet Trop Dis, ZA-0110 Onderstepoort, South Africa
[2] Univ Pretoria, Equine Res Ctr, ZA-0110 Onderstepoort, South Africa
[3] AFRC, Inst Anim Hlth, Pirbright Lab, Woking GU24 0NF, Surrey, England
关键词
equine encephalosis virus; EEV; sandwich ELISA; group-specific; polyclonal antibody;
D O I
10.1016/S0166-0934(03)00202-7
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A polyclonal antibody-based, group-specific, indirect, sandwich ELISA (S-ELISA) for the detection of equine encephalosis virus (EEV) antigen was developed. Purified EEV particles were titrated in the S-ELISA and the limit of detection was determined to be approximately 9.0 ng of antigen/ml (0.45 ng/well). Positive S-ELISA reactions were recorded with seven serologically distinct EEV serotypes. No cross-reactions were recorded with other arboviruses including African horse sickness virus (AHSV) serotypes 1-9, bluetongue serotypes 1-24, epizootic haemorrhagic disease serotypes 1-8 and isolate 318, and selected isolates of Palyam, Eubenangee, Corriparta, Warrego, Akabane and bovine ephemeral fever viruses. The assay proved to be sensitive and specific for the rapid detection of EEV in cell cultures and in homogenated suckling mouse brain (MB). The data generated in this study suggest that the ELISA will be valuable for epidemiological studies of EE and will assist in making a reliable differential diagnosis between EEV and AHSV infections. (C) 2003 Elsevier B.V. All rights reserved.
引用
收藏
页码:129 / 135
页数:7
相关论文
共 12 条
  • [1] [Anonymous], VIRUS TAXONOMY 7 REP
  • [2] Barnard BJH, 1997, ONDERSTEPOORT J VET, V64, P95
  • [3] BURR GW, 1994, OSA TECHNICAL DIGEST, V7, P9
  • [4] COETZER JAW, 1994, INFECT DIS LIVESTOCK, P476
  • [5] CRAFFORD JE, 2002, THESIS U PRETORIA
  • [6] A SEROGROUP SPECIFIC ENZYME-LINKED-IMMUNOSORBENT-ASSAY FOR THE DETECTION AND IDENTIFICATION OF AFRICAN HORSE SICKNESS VIRUSES
    HAMBLIN, C
    MERTENS, PPC
    MELLOR, PS
    BURROUGHS, JN
    CROWTHER, JR
    [J]. JOURNAL OF VIROLOGICAL METHODS, 1991, 31 (2-3) : 285 - 292
  • [7] Howell PG, 2002, ONDERSTEPOORT J VET, V69, P79
  • [8] Karber G., 1931, ARCH EXP PATH PHARMA, V162, P480, DOI [DOI 10.1007/BF01863914, 10.1007/BF01863914]
  • [9] PURIFICATION AND PROPERTIES OF VIRUS-PARTICLES, INFECTIOUS SUBVIRAL PARTICLES, AND CORES OF BLUETONGUE VIRUS SEROTYPE-1 AND SEROTYPE-4
    MERTENS, PPC
    BURROUGHS, JN
    ANDERSON, J
    [J]. VIROLOGY, 1987, 157 (02) : 375 - 386
  • [10] PAWESKA JT, 1999, EQUINE INFECT DIS, V8, P303