ABSTRACT Rathayibacter toxicus is Gram-positive bacterium causing gummosis of forage grasses, and it produces a corynetoxin responsible for livestock poisonings in Australia. R. toxicus is a USDA APHIS-listed Select Agent, threatening seed and livestock production areas in the U.S. We therefore are developing diagnostic assays to prepare for the introduction of the pathogen into the U.S. Using proteomic data, we identified proteins expressed in R. toxicus but not in other Rathayibacter spp. Recombinant proteins corresponding to extracellular domains of candidate proteins were used to raise polyclonal and monoclonal antibodies. Antibodies were evaluated for sensitivity and specificity in immunoassays against a panel of Rathayibacter spp. and near-neighbours. Polyclonal and monoclonal antibodies exhibited high sensitivity and specificity for R. toxicus in assays with cultured bacteria. The highest-performing immunoreagents were identified for further testing with field samples collected in Australia and for eventual development of an immunoassay for bacterial detection in grass samples.

中文翻译：

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用于开发特定于毒性 Rathayibacter 的诊断测定的免疫试剂

摘要 Rathayibactertoxicus 是革兰氏阳性菌，会引起牧草的胶质病，它会产生一种棒状毒素，导致澳大利亚的牲畜中毒。R.toxicus 是美国农业部 APHIS 列出的选择剂，威胁着美国的种子和牲畜生产区 我们因此正在开发诊断分析，为将病原体引入美国做准备 使用蛋白质组学数据，我们鉴定了 R.toxicus 中表达的蛋白质但不是在其他 Rathayibacter spp。对应于候选蛋白胞外域的重组蛋白被用于产生多克隆和单克隆抗体。在针对一组 Rathayibacter spp 的免疫测定中评估了抗体的敏感性和特异性。和近邻。多克隆和单克隆抗体对 R 表现出高灵敏度和特异性。培养细菌检测中的毒性。性能最高的免疫试剂被确定用于进一步测试在澳大利亚收集的现场样品，并最终开发用于草样品中细菌检测的免疫测定法。