Infantes-Lorenzo Jose A, Dave Dipesh, Moreno Immaculada, Anderson Paul, Lesellier Sandrine, Gormley Eamonn, Dominguez Lucas, Balseiro Ana, Gortázar Christian, Dominguez Mercedes, Salguero Francisco J
VISAVET Health Surveillance Centre, Universidad Complutense de Madrid, Madrid, Spain.
Bacteriology Department, Animal and Plant Health Agency, Addlestone, Surrey, UK.
Vet Med Sci. 2019 Feb;5(1):61-69. doi: 10.1002/vms3.134. Epub 2019 Jan 18.
European badgers (Meles meles) have been identified as wildlife reservoirs for Mycobacterium bovis in the UK and Ireland, and may also have a role in the epidemiology of animal tuberculosis in other European regions. Thus, detection of M. bovis-infected badgers may be required for the purposes of surveillance and monitoring of disease levels in infected populations. Current serological assays to detect M. bovis infection in live badgers, while rapid and inexpensive, show limited diagnostic sensitivity. Here we describe and evaluate new ELISA platforms for the recognition of the P22 multiprotein complex derived from the purified protein derivative (PPD) of M. bovis. The recognition of IgG against P22 multiprotein complex derived from PPD-B was tested by ELISA in the serum of badgers from the UK, Ireland and Spain. TB infection in the badgers was indicated by the presence of M. bovis in tissues by culture and histology at post-mortem examination and TB-free status was established by repeated negativity in the interferon γ release assay (IGRA). In experimentally infected badgers, humoral antibody responses against P22 developed within 45 days post-infection. The ELISA tests showed estimated sensitivity levels of 74-82% in experimentally and naturally infected badgers with specificities ranging from 75% to 100% depending on the badger population tested. The P22 multi-antigen based ELISAs provide a sensitive and specific test platform for improved tuberculosis surveillance in badgers.
在英国和爱尔兰，欧洲獾（Meles meles）已被确定为牛分枝杆菌的野生动物宿主，并且在其他欧洲地区的动物结核病流行病学中可能也起到一定作用。因此，为了监测感染种群中的疾病水平，可能需要检测感染牛分枝杆菌的獾。目前用于检测活体獾牛分枝杆菌感染的血清学检测方法虽然快速且成本低廉，但诊断敏感性有限。在此，我们描述并评估了用于识别源自牛分枝杆菌纯化蛋白衍生物（PPD）的P22多蛋白复合物的新型ELISA平台。通过ELISA检测了来自英国、爱尔兰和西班牙的獾血清中针对源自PPD-B的P22多蛋白复合物的IgG识别情况。獾的结核病感染通过死后组织培养和组织学检查中牛分枝杆菌的存在来表明，而通过干扰素γ释放试验（IGRA）反复呈阴性来确定无结核病状态。在实验感染的獾中，感染后45天内出现了针对P22的体液抗体反应。ELISA检测在实验感染和自然感染的獾中显示出估计的敏感性水平为74%-82%，特异性根据所检测的獾种群不同，范围为75%至100%。基于P22多抗原的ELISA为改进獾结核病监测提供了一个敏感且特异的检测平台。
