Kökkaya Serkan, Göksu Ayşe Gençay
Department of Microbiology, Faculty of Veterinary Medicine, Yozgat Bozok University, Yozgat, Türkiye.
Department of Virology, Faculty of Veterinary Medicine, Erciyes University, Kayseri, Türkiye.
Vet Med Sci. 2025 Sep;11(5):e70516. doi: 10.1002/vms3.70516.
Canine distemper virus (CDV) is a highly contagious pathogen that affects several animal families. While animals of all ages are susceptible, puppies face the highest risk. This study aimed to detect CDV in dogs with suspected infection, isolate the virus and develop a sensitive antigen-ELISA test using monoclonal and polyclonal antibodies. Samples were collected from 40 dogs and tested by RT-PCR, then cultured positive samples in MDCK cells for virus isolation. In addition, the CDV F protein gene sequence obtained from the MDCK-34388 isolate was added to GenBank. Phylogenetic analysis revealed that the isolate belongs to the Asia-1 genotype. Among the 40 tested dogs, 29 (72.5%) tested positive for CDV, exhibiting respiratory, neurological and mixed symptoms in 62.5%, 15% and 22.5% of cases, respectively. Using an inactivated CDV isolate, we produced monoclonal antibodies in BALB/c mice and polyclonal antibodies in New Zealand rabbits. For ELISA development, we HRP-labelled the 1G10-1A monoclonal antibody that targets the CDV F protein. The antigen ELISA test we developed showed 90% sensitivity and 87.5% specificity, with positive and negative predictive values of 87.8% and 89.74%, respectively. These findings indicate that the antigen ELISA test is a reliable diagnostic tool for CDV detection, complementing RT-PCR and supporting effective disease management for veterinary practice.
犬瘟热病毒（CDV）是一种极具传染性的病原体，可感染多个动物科。虽然所有年龄段的动物都易感染，但幼犬面临的风险最高。本研究旨在检测疑似感染犬瘟热病毒的犬只，分离病毒，并使用单克隆抗体和多克隆抗体开发一种灵敏的抗原酶联免疫吸附测定（ELISA）检测方法。从40只犬采集样本并通过逆转录聚合酶链反应（RT-PCR）进行检测，然后将阳性样本在犬肾传代细胞（MDCK）中培养以进行病毒分离。此外，从MDCK-34388分离株获得的犬瘟热病毒融合蛋白（F）基因序列被添加到基因库中。系统发育分析表明，该分离株属于亚洲-1基因型。在40只受试犬中，29只（72.5%）犬瘟热病毒检测呈阳性，分别有62.5%、15%和22.5%的病例表现出呼吸道、神经和混合症状。我们使用一株灭活的犬瘟热病毒分离株，在BALB/c小鼠中制备了单克隆抗体，在新西兰兔中制备了多克隆抗体。为开发ELISA检测方法，我们用辣根过氧化物酶（HRP）标记了靶向犬瘟热病毒F蛋白的1G10-1A单克隆抗体。我们开发的抗原ELISA检测方法灵敏度为90%，特异性为87.5%，阳性预测值和阴性预测值分别为87.8%和89.74%。这些研究结果表明，抗原ELISA检测方法是一种可靠的犬瘟热病毒检测诊断工具，可作为RT-PCR的补充，并为兽医实践中的有效疾病管理提供支持。
