Muller Janine D, McEachern Jennifer A, Bossart Katharine N, Hansson Eric, Yu Meng, Clavijo Alfonso, Hammond Jef M, Wang Lin-Fa
CSIRO Livestock Industries, Australian Animal Health Laboratory, 5 Portarlington Road, Geelong, Victoria 3220, Australia.
J Virol Methods. 2008 Jul;151(1):146-53. doi: 10.1016/j.jviromet.2008.03.011. Epub 2008 Apr 25.
Foot-and-mouth disease virus (FMDV) causes a highly contagious vesicular disease affecting cloven hoofed animals and is considered the most economically important disease worldwide. Recent FMD outbreaks in Europe and Taiwan and the associated need for rapid diagnostic turnaround have identified limitations that exist in current diagnostic capabilities. To aid improved diagnosis, a serotype-independent FMDV antigen capture assay was developed using antibodies directed against a highly conserved cross-reactive protein fragment (1AB') located within the structural protein 1AB. Cattle sera raised against all 7 serotypes of FMDV bound purified 1AB' demonstrating its immunogenicity in infected animals. Polyclonal anti-1AB' antiserum was produced in chickens and applied as a universal detector of FMDV antigen. Western blot analysis and ELISA both demonstrated that anti-1AB' serum could recognize FMDV antigens independent of serotype. Two recently characterized anti-FMDV monoclonal antibodies were also evaluated for their ability to capture FMDV antigen independently of serotype. When used in combination with chicken anti-1AB' antibodies in an antigen capture ELISA format, all serotypes of FMDV were detected. These data represent the first demonstration of the use of serotype-independent FMDV antigen capture reagents which may enable the development of rapid laboratory based assays or perhaps more significantly, rapid field-based pen-side or point of entry border control diagnostic tests.
口蹄疫病毒(FMDV)引发一种极具传染性的水疱病,影响偶蹄类动物,被认为是全球经济上最重要的疾病。近期在欧洲和台湾地区爆发的口蹄疫疫情,以及随之而来的对快速诊断周转的需求,凸显了当前诊断能力存在的局限性。为了助力改进诊断,利用针对位于结构蛋白1AB内高度保守的交叉反应蛋白片段(1AB')的抗体,开发了一种与血清型无关的口蹄疫病毒抗原捕获检测方法。用针对口蹄疫病毒所有7种血清型产生的牛血清与纯化的1AB'结合,证明了其在感染动物中的免疫原性。在鸡中制备了多克隆抗1AB'抗血清,并将其用作口蹄疫病毒抗原的通用检测剂。蛋白质印迹分析和酶联免疫吸附测定均表明,抗1AB'血清能够识别与血清型无关的口蹄疫病毒抗原。还评估了两种最近鉴定的抗口蹄疫病毒单克隆抗体独立于血清型捕获口蹄疫病毒抗原的能力。当在抗原捕获酶联免疫吸附测定形式中与鸡抗1AB'抗体联合使用时,能够检测到口蹄疫病毒的所有血清型。这些数据首次证明了使用与血清型无关的口蹄疫病毒抗原捕获试剂,这可能有助于开发基于实验室的快速检测方法,或者更重要的是,开发基于现场的畜栏旁或入境口岸边境控制快速诊断测试。