Tian Yutong, Zhang Hailong, Zhang Yan, Zhang Xinya, Guan Zhilei, Zhang Junjie, Qiu Yafeng, Li Beibei, Liu Ke, Li Zongjie, Shao Donghua, Li Peng, Ma Zhiyong, Wei Jianchao
College of Animal Science, Yangtze University, Jingzhou 434025, China.
Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China.
Vet Sci. 2024 Mar 21;11(3):138. doi: 10.3390/vetsci11030138.
Caprine arthritis encephalitis is an infectious disease caused by the caprine arthritis encephalitis virus that infects goats, sheep, and other small ruminants. An outbreak of CAEV could be extremely harmful to the goat farming industry and could cause severe economic losses. We designed specific primers and probes for the gag gene and established a TaqMan real-time quantitative polymerase chain reaction assay. This method's correlation coefficient (R) was >0.999, and the sensitivity of the assay to the plasmid-carried partial gag gene was approximately 10 copies/µL, 1000 times higher than that of conventional PCR. No specific fluorescence was detected for other sheep viruses. Using this method, we tested 776 asymptomatic sheep blood samples and 4 neurodegenerative sheep brain samples from six farms in eastern China, and the positivity rate was 0.77% (6/780). The gag gene was partially sequenced in the three positive samples and compared with the sequences from other representative strains in GenBank. The results revealed that all three strains belonged to the B1 subtype and were most closely related to the strains from Shanxi and Gansu, previously isolated in China, with their homology ranging from 97.7% to 98.9%. These results suggest that the designed RT-qPCR assay can be used to detect subclinical CAEV in sheep and that the virus is still present in eastern China.
山羊关节炎脑炎是由山羊关节炎脑炎病毒引起的一种传染病，该病毒可感染山羊、绵羊和其他小型反刍动物。山羊关节炎脑炎病毒的爆发可能对山羊养殖业造成极大危害，并可能导致严重的经济损失。我们针对gag基因设计了特异性引物和探针，并建立了TaqMan实时定量聚合酶链反应检测方法。该方法的相关系数（R）>0.999，对携带质粒的部分gag基因的检测灵敏度约为10拷贝/µL，比传统PCR高1000倍。对其他绵羊病毒未检测到特异性荧光。我们使用该方法对来自中国东部六个养殖场的776份无症状绵羊血液样本和4份神经退行性变绵羊脑样本进行了检测，阳性率为0.77%（6/780）。对三个阳性样本中的gag基因进行了部分测序，并与GenBank中其他代表性菌株的序列进行了比较。结果显示，所有三个菌株均属于B1亚型，与中国此前在山西和甘肃分离的菌株关系最为密切，同源性在97.7%至98.9%之间。这些结果表明，所设计的逆转录定量聚合酶链反应检测方法可用于检测绵羊亚临床感染的山羊关节炎脑炎病毒，且该病毒在中国东部地区仍然存在。
