Desport Moira, Stewart Meredith E, Sheridan Carol A, Ditcham William G F, Setiyaningsih Surachmi, Tenaya W Masa, Hartaningsih Nining, Wilcox Graham E
Department of Veterinary and Biomedical Sciences, Murdoch University, Murdoch, WA 6150, Australia.
J Virol Methods. 2005 Mar;124(1-2):135-42. doi: 10.1016/j.jviromet.2004.11.015. Epub 2004 Dec 29.
In Indonesia, it is suspected that there are two bovine lentiviruses circulating in the cattle population: a pathogenic Jembrana disease virus (JDV), and a nonpathogenic bovine immunodeficiency-like virus (BIV). Both viruses cross-react antigenically and cannot be differentiated by current serological tests using JDV antigens. To identify possible type-specific epitopes, a series of recombinant protein constructs including the matrix, capsid and nucleocapsid proteins were produced from JDV gag and the expressed proteins were tested by Western blot using JDV and BIV hyperimmune sera. JDV matrix and truncated capsid proteins were recognised by both JDV and BIV hyperimmune sera indicating that there were multiple cross-reactive epitopes present in JDV gag. At least three epitopic regions were identified in these constructs, including the major homology region, by monoclonal antibody binding studies. JDV nucleocapsid recombinant protein was not recognised by either JDV or BIV hyperimmune sera and none of the recombinant gag proteins were able to differentiate between JDV positive sera from Jembrana disease endemic and Jembrana disease-free areas. Additionally, a 40 amino acid recombinant subunit protein encompassing the region recently found to contain an epitope unique to BIV [Zheng, L., Zhang, S., Wood, C., Kapil, S., Wilcox, G.E., Loughin, T.A., Minocha, H.C., 2001. Differentiation of two bovine lentiviruses by a monoclonal antibody on the basis of epitope specificity. Clin. Diagn. Lab. Immunol. 8, 283-287] was tested but was not recognised by either JDV positive sera from Jembrana disease-endemic or Jembrana disease-free areas.
在印度尼西亚,人们怀疑有两种牛慢病毒在牛群中传播:致病性的杰姆纳纳病病毒(JDV)和非致病性的牛免疫缺陷样病毒(BIV)。这两种病毒在抗原上会发生交叉反应,无法通过目前使用JDV抗原的血清学检测进行区分。为了确定可能的型特异性表位,从JDV gag基因产生了一系列重组蛋白构建体,包括基质蛋白、衣壳蛋白和核衣壳蛋白,并使用JDV和BIV超免疫血清通过蛋白质印迹法对表达的蛋白进行检测。JDV基质蛋白和截短的衣壳蛋白可被JDV和BIV超免疫血清识别,这表明JDV gag基因中存在多个交叉反应表位。通过单克隆抗体结合研究,在这些构建体中至少鉴定出三个表位区域,包括主要同源区域。JDV核衣壳重组蛋白未被JDV或BIV超免疫血清识别,并且没有一种重组gag蛋白能够区分来自杰姆纳纳病流行地区和无杰姆纳纳病地区的JDV阳性血清。此外,还检测了一种包含最近发现的含有BIV独特表位区域的40个氨基酸的重组亚基蛋白[郑,L.,张,S.,伍德,C.,卡皮尔,S.,威尔科克斯,G.E.,洛金,T.A.,米诺查,H.C.,2001年。基于表位特异性的单克隆抗体对两种牛慢病毒的区分。临床诊断实验室免疫学。8,283 - 287],但它未被来自杰姆纳纳病流行地区或无杰姆纳纳病地区的JDV阳性血清识别。