利用重组衣壳蛋白和跨膜蛋白检测羊慢病毒抗体。

V. Celer, V. Celer
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引用次数: 9

摘要

采用聚合酶链反应扩增maedii - visna病毒衣壳蛋白p25和跨膜蛋白的编码序列,并将其克隆到质粒表达载体pRSET-B中。这两种DNA结构都表达了带有多组氨酸标记的蛋白质。用Ni-NTA琼脂糖纯化重组蛋白,并用免疫印迹法检测maedii - visna病毒抗体。对260份羊血清标本进行了分析。p25阳性血清111份(42.7%)。rTM抗原具有较高的敏感性,在118份(45.4%)血清中检测到抗体。两种重组蛋白结合作为抗原的血清学检测灵敏度高于整个病毒抗原。
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Detection of antibodies to ovine lentivirus using recombinant capsid and transmembrane proteins.
The coding sequences of the capsid protein p25 and transmembrane protein of Maedi-Visna virus were amplified using polymerase chain reaction and cloned into the plasmid expression vector pRSET-B. Both DNA constructs expressed proteins tagged with polyhistidine. The recombinant proteins were purified using Ni-NTA agarose and used in immunoblot to detect antibodies against Maedi-Visna virus. A total of 260 ovine serum specimens was analysed. The total number of p25-positive sera was 111 (42.7%). Higher sensitivity was achieved with rTM antigen, which detected antibodies in 118 (45.4%) sera. The combination of both recombinant proteins as antigens resulted in higher sensitivity of serological detection compared to whole virus antigen.
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