Dalley Deanna, Lesellier Sandrine, Salguero Francisco J, Chambers Mark A
Department of Bacteriology, Animal and Plant Health Agency, Addlestone, Surrey KT15 3NB, UK.
Laboratoire de la Rage et de la Faune Sauvage de Nancy (LRFSN), ANSES, 54220 Malzéville, France.
Vet Sci. 2019 Nov 2;6(4):89. doi: 10.3390/vetsci6040089.
European badgers are a wildlife reservoir of bovine tuberculosis in parts of Great Britain. Accurate diagnosis of tuberculosis in badgers is important for the development of strategies for the control of the disease. Sensitive serological tests for badger TB are needed for reasons such as cost and simplicity. Assay of mucosal IgA could be useful for diagnosing respiratory pathogens such as and for monitoring the response to mucosal vaccination. To develop an IgA assay, we purified secretory IgA from badger bile, identifying secretory component (SC), heavy chain (HC) and light chain (LC), at 66, 46 and 27 Kda, respectively, on the basis of size comparison with other species. Monoclonal antibodies (mAbs) were generated to purified IgA. We selected two for ELISA development. The detection limit of the IgA-specific mAbs was found to be approximately 20 ng/mL when titrated against purified badger bile. One monoclonal antibody specific for badger IgA was used to detect IgA in serum and tracheal aspirate with specificity to an immunodominant antigen of . An infection dose-dependent IgA response was observed in experimentally infected badgers. IgA was also detected by immunohistochemistry in the lungs of bTB-infected badgers. With further characterisation, these represent new reagents for the study of the IgA response in badgers.
在英国部分地区，欧洲獾是牛结核病的野生动物宿主。准确诊断獾的结核病对于制定疾病控制策略至关重要。出于成本和简便性等原因，需要对獾结核病进行敏感的血清学检测。检测黏膜IgA可能有助于诊断诸如[此处原文缺失病原体名称]等呼吸道病原体，并监测对黏膜疫苗接种的反应。为了开发一种IgA检测方法，我们从獾胆汁中纯化了分泌型IgA，通过与其他物种的大小比较，分别鉴定出分子量为66 kDa的分泌成分（SC）、46 kDa的重链（HC）和27 kDa的轻链（LC）。针对纯化的IgA产生了单克隆抗体（mAb）。我们选择了两种用于酶联免疫吸附测定（ELISA）的开发。当用纯化的獾胆汁进行滴定检测时，发现IgA特异性单克隆抗体的检测限约为20 ng/mL。一种对獾IgA特异的单克隆抗体被用于检测血清和气管吸出物中的IgA，该IgA对[此处原文缺失病原体名称]的一种免疫显性抗原有特异性。在实验感染的獾中观察到了感染剂量依赖性的IgA反应。通过免疫组织化学也在感染牛结核病的獾的肺中检测到了IgA。随着进一步的特性鉴定，这些代表了用于研究獾中IgA反应的新试剂。
