HIGHLY SPECIFIC CONFIRMATORY WESTERN-BLOT TEST FOR AFRICAN SWINE FEVER VIRUS-ANTIBODY DETECTION USING THE RECOMBINANT VIRUS PROTEIN P54
HIGHLY SPECIFIC CONFIRMATORY WESTERN-BLOT TEST FOR AFRICAN SWINE FEVER VIRUS-ANTIBODY DETECTION USING THE RECOMBINANT VIRUS PROTEIN P54
复制标题
DOI：
10.1016/0166-0934(94)00150-f
复制
发表时间：
1995-03-01
影响因子：
3.1
通讯作者：
ESCRIBANO, JM
中科院分区：
文献类型：
--
作者：
ALCARAZ, C;RODRIGUEZ, F;ESCRIBANO, JM
建立了一种使用重组蛋白的Western印迹技术，以确认用ELISA检测非洲猪瘟(ASF)特异性抗体的阳性结果。新的确认性Western印迹是基于蛋白P54的使用，P54是最具抗原性的ASF病毒结构蛋白之一，在大肠杆菌中表达，与MS2聚合酶的N端融合。重组Western印迹方法对ASF病毒感染猪的检测具有高度的特异性和同样的敏感性，与传统的Western印迹方法一样，后者使用的是病毒诱导的蛋白质，分子量在23到35 kDa之间。这种新颖的Western印迹分析方法对测试提供了更简单的解释，消除了传统技术中使用的污染抗原的细胞化合物产生假阳性反应的可能性，并避免了在抗原生产中使用活病毒。
A Western blot technique using a recombinant protein has been developed to confirm positive results obtained in African swine fever (ASF)-specific antibody detection by ELISA. The new confirmatory Western blot is based on the use of protein p54, one of the most antigenic ASF virus structural proteins, expressed in Escherichia coli fused to the N-terminus of MS2 polymerase. The recombinant Western blot assay was highly specific and equally sensitive for ASF virus-infected pigs detection as the conventional Western blot, which uses virus-induced proteins ranging in molecular weight between 23 and 35 kDa. The novel Western blot assay provides a simpler interpretation of the test, eliminates the possibility of false-positive reactions produced by cellular compounds that contaminate the antigen employed in the conventional technique, and avoids the use of live virus in antigen production.