Liu Fengli, Cao Yanxin, Yan Maokai, Sun Mengxu, Zhang Qingshui, Wang Jun, Fu Guanghua, Liu Rongchang, Huang Yu, Su Jingliang
Key Laboratory of Animal Epidemiology of the Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China.
Agricultural and Environmental Branch, Jiaxing Vocational and Technical College, Jiaxing 314036, China.
Pathogens. 2021 Mar 18;10(3):365. doi: 10.3390/pathogens10030365.
Duck viral enteritis is a highly contagious and fatal disease of commercial waterfowl flocks. The disease occurs sporadically or epizootically in mainland China due to insufficient vaccinations. Early and rapid diagnosis is important for preventive intervention and the control of epizootic events in clinical settings. In this study, we generated two monoclonal antibodies (MAbs) that specifically recognized the duck enteritis virus (DEV) envelope glycoprotein B and tegument protein UL47, respectively. Using these MAbs, a colloidal gold-based immunochromatographic assay (ICA) was developed for the efficient detection of DEV antigens within 15 min. Our results showed that the detection limit of the developed ICA strip was 2.52 × 10 TCID/mL for the virus infected cell culture suspension with no cross-reactivity with other pathogenic viruses commonly encountered in commercially raised waterfowl. Using samples from experimentally infected ducks, we demonstrated that the ICA detected the virus in cloacal swab samples on day three post-infection, demonstrating an 80% concordance with the PCR. For tissue homogenates from ducks succumbing to infection, the detection sensitivity was 100%. The efficient and specific detection by this ICA test provides a valuable, convenient, easy to use and rapid diagnostic tool for DVE under both laboratory and field conditions.
鸭病毒性肠炎是一种对商业水禽群具有高度传染性和致命性的疾病。由于疫苗接种不足，该病在中国内地呈散发性或流行发病。早期快速诊断对于临床环境中的预防干预和疫情控制至关重要。在本研究中，我们分别制备了两种特异性识别鸭肠炎病毒（DEV）包膜糖蛋白B和衣壳蛋白UL47的单克隆抗体（MAb）。利用这些单克隆抗体，开发了一种基于胶体金的免疫层析检测法（ICA），可在15分钟内高效检测DEV抗原。我们的结果表明，所开发的ICA试纸条对病毒感染细胞培养悬液的检测限为2.52×10 TCID/mL，与商业养殖水禽中常见的其他致病病毒无交叉反应。使用来自实验感染鸭的样本，我们证明ICA在感染后第三天在泄殖腔拭子样本中检测到了病毒，与PCR的一致性为80%。对于死于感染的鸭的组织匀浆，检测灵敏度为100%。这种ICA检测方法的高效特异性检测为实验室和现场条件下的鸭病毒性肠炎提供了一种有价值、方便、易用且快速的诊断工具。
