Key Laboratory of Jiangsu Preventive Veterinary Medicine, Key Laboratory for Avian Preventive Medicine, Ministry of Education, College of Veterinary Medicine, Yangzhou University, Yangzhou, 225009, Jiangsu, China.
Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, 225009, Jiangsu, China.
BMC Vet Res. 2019 Jul 8;15(1):232. doi: 10.1186/s12917-019-1987-5.
Recently, serotype 4 fowl adenovirus (FAdV-4) has spread widely and caused huge economic loss to poultry industry. However, little is known about the molecular pathogenesis of FAdV-4. Fiber protein is thought to be vital for its infection and pathogenesis.
Two novel monoclonal antibodies (mAbs) targeting the fiber-1 protein of FAdV-4 were generated, designated as mAb 3B5 and 6H9 respectively. Indirect immunofluorescence assay (IFA) showed that both mAbs only reacted with the FAdV-4 and FAdV-10, not with other serotypes including FAdV-1, FAdV-5, FAdV-6, FAdV-7, FAdV-8 and FAdV-9 tested. Although both mAbs did not recognize the linear epitopes, they could efficiently immunoprecipitate the fiber-1 protein in LMH cells either infected with FAdV-4 or transfected with pcDNA3.1-Fiber-1. Moreover, mAb 3B5 as a capture antibody and HRP-conjugated mAb 6H9 as a detection antibody, a novel sandwich ELISA for efficient detection of FAdV-4 was generated. The limit of detection of the ELISA could reach to 1000 TCID/ml of FAdV-4 and the ELISA could be efficiently applied to detect FAdV-4 in the clinical samples.
The two mAbs specific targeting fiber-1 generated here would pave the way for further studying on the role of fiber-1 in the infection and pathogenesis of FAdV-4, and the established mAb based sandwich ELISA would provide an efficient diagnostics tool for detection of FAdV-4/10.
最近，血清 4 型禽腺病毒（FAdV-4）广泛传播，给家禽业造成了巨大的经济损失。然而，人们对 FAdV-4 的分子发病机制知之甚少。纤维蛋白被认为对其感染和发病机制至关重要。
生成了两种针对 FAdV-4 纤维蛋白-1 的新型单克隆抗体（mAb），分别命名为 mAb 3B5 和 6H9。间接免疫荧光试验（IFA）表明，这两种 mAb 仅与 FAdV-4 和 FAdV-10 反应，而与其他血清型（包括 FAdV-1、FAdV-5、FAdV-6、FAdV-7、FAdV-8 和 FAdV-9）不反应。尽管这两种 mAb 均不识别线性表位，但它们都能有效地免疫沉淀在 LMH 细胞中感染 FAdV-4 或转染 pcDNA3.1-Fiber-1 的纤维-1 蛋白。此外，mAb 3B5 作为捕获抗体和 HRP 标记的 mAb 6H9 作为检测抗体，建立了一种用于高效检测 FAdV-4 的新型夹心 ELISA。该 ELISA 的检测限可达到 1000 TCID/ml 的 FAdV-4，并且该 ELISA 可有效地用于检测临床样本中的 FAdV-4。
这里生成的两种针对纤维-1 的特异性 mAb 将为进一步研究纤维-1 在 FAdV-4 感染和发病机制中的作用铺平道路，建立的基于 mAb 的夹心 ELISA 将为 FAdV-4/10 的检测提供一种高效的诊断工具。
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