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一种利用重组融合蛋白检测马动脉炎病毒宿主抗体存在的酶联免疫吸附测定法的开发与评估。

Development and evaluation of an ELISA using recombinant fusion protein to detect the presence of host antibody to equine arteritis virus.

作者信息

Chirnside E D, Francis P M, de Vries A A, Sinclair R, Mumford J A

机构信息

Department of Infectious Diseases, Animal Health Trust, Lanwades Park, Kennett, Newmarket, Suffolk, UK.

出版信息

J Virol Methods. 1995 Jul;54(1):1-13. doi: 10.1016/0166-0934(95)00020-u.

Abstract

A recombinant glutathione-S-transferase fusion protein expressing amino acids 55-98 of equine arteritis virus (EAV) GL (rGL 55-98) was tested in an ELISA for its ability to detect serum antibodies to EAV. Host antibodies induced following EAV infection bound the recombinant antigen by ELISA. The ELISA specificity and sensitivity were determined with a panel of equine sera including postinfection and postvaccination samples. A good correlation existed between EAV neutralizing antibody titers and ELISA absorbance values (r = 0.827). The sensitivity and specificity of the ELISA were 99.6 and 90.1%, respectively, compared with EAV neutralization test and the recombinant antigen did not crossreact in ELISA with equine sera directed against other common equine respiratory viruses. Three post-EAV infection equine sera raised against different EAV isolates reacted strongly in the ELISA, as did two equine sera raised against EAV vaccines, indicating that the viral epitope was conserved between the viruses tested. Following vaccination with an inactivated whole virus vaccine, antibody detected with the recombinant antigen ELISA preceded the development of a virus-neutralizing response. The study demonstrates the potential application of rGL 55-98 as a diagnostic antigen.

摘要

对一种表达马动脉炎病毒(EAV)GL蛋白55 - 98位氨基酸的重组谷胱甘肽-S-转移酶融合蛋白(rGL 55 - 98)进行了酶联免疫吸附测定(ELISA),以检测其检测抗EAV血清抗体的能力。EAV感染后诱导产生的宿主抗体通过ELISA与重组抗原结合。用一组马血清(包括感染后和接种疫苗后的样本)测定了ELISA的特异性和敏感性。EAV中和抗体滴度与ELISA吸光度值之间存在良好的相关性(r = 0.827)。与EAV中和试验相比,ELISA的敏感性和特异性分别为99.6%和90.1%,并且该重组抗原在ELISA中与针对其他常见马呼吸道病毒的马血清无交叉反应。三份针对不同EAV分离株产生的EAV感染后马血清在ELISA中反应强烈,两份针对EAV疫苗产生的马血清也是如此,这表明所测试病毒之间的病毒表位是保守的。在用灭活全病毒疫苗接种后,用重组抗原ELISA检测到的抗体先于病毒中和反应出现。该研究证明了rGL 55 - 98作为诊断抗原的潜在应用。

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