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非洲猪瘟病毒主要抗原蛋白p54和p30在杆状病毒中的高水平表达及其作为诊断试剂的潜在用途。

High level expression of the major antigenic African swine fever virus proteins p54 and p30 in baculovirus and their potential use as diagnostic reagents.

作者信息

Oviedo J M, Rodríguez F, Gómez-Puertas P, Brun A, Gómez N, Alonso C, Escribano J M

机构信息

Centro de Investigación en Sanidad Animal (CISA-INIA), Madrid, Spain.

出版信息

J Virol Methods. 1997 Feb;64(1):27-35. doi: 10.1016/s0166-0934(96)02140-4.

Abstract

At present, the eradication of African swine fever (ASF) in affected countries is based only on an efficient diagnosis program because of the absence of an available vaccine. The highly antigenic ASF virus proteins p54 and p30, encoded by genes E183L and CP204L respectively, were expressed in baculovirus for diagnostic purposes. A sequence comparison analysis of these genes from different field virus strains which are geographically diverse and isolated in different years, revealed that both genes are completely conserved among the isolates. Partially purified baculovirus-expressed proteins were used in ELISA and Western blot for ASF antibody detection in sera from experimentally inoculated pigs and field sera from ASF innaparent carriers. These comparative analyses showed that p54 presents better reactivity than p30 in Western blot. However, recombinant p30 was more efficient for antibody detection by ELISA, improving the discrimination between positive and negative sera by this technique. These data suggest the convenience of using p30 as ELISA antigen, while p54 should be the selected antigen for ASF virus antibody detection by Western blot. The combined use of both antigens for serodiagnosis of ASF disease will improve the sensitivity of innaparent carriers detection, facilitating also the interpretation of the tests, and eliminating the use of ASF virus in antigen production.

摘要

目前,由于缺乏可用疫苗,受影响国家的非洲猪瘟(ASF)根除工作仅基于高效的诊断程序。分别由基因E183L和CP204L编码的高抗原性ASF病毒蛋白p54和p30在杆状病毒中表达用于诊断目的。对来自不同地理区域且在不同年份分离的不同野外病毒株的这些基因进行序列比较分析,结果显示这两个基因在分离株中完全保守。部分纯化的杆状病毒表达蛋白用于ELISA和Western印迹,以检测实验接种猪的血清和ASF隐性携带者的野外血清中的ASF抗体。这些比较分析表明,在Western印迹中p54比p30表现出更好的反应性。然而,重组p30在ELISA抗体检测中效率更高,通过该技术提高了阳性和阴性血清之间的区分度。这些数据表明使用p30作为ELISA抗原的便利性,而p54应作为Western印迹检测ASF病毒抗体的选定抗原。联合使用这两种抗原进行ASF疾病的血清学诊断将提高隐性携带者检测的灵敏度,也便于试验结果的解读,并消除在抗原生产中使用ASF病毒。

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