Detection and Phylogenetic Analysis of Caprine Arthritis Encephalitis Virus Using TaqMan-based qPCR in Eastern China

文献类型： 外文期刊

第一作者： Tian, Yutong

作者： Tian, Yutong;Li, Peng;Tian, Yutong;Zhang, Hailong;Zhang, Yan;Zhang, Xinya;Guan, Zhilei;Zhang, Junjie;Qiu, Yafeng;Li, Beibei;Liu, Ke;Li, Zongjie;Shao, Donghua;Ma, Zhiyong;Wei, Jianchao;Zhang, Hailong;Zhang, Yan

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关键词： caprine arthritis encephalitis virus; TaqMan RT-qPCR; goat; phylogenetic trees; detecting method; infectious diseases

期刊名称：VETERINARY SCIENCES （ 影响因子：2.4； ）

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年卷期： 2024 年 11 卷 3 期

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收录情况： SCI

摘要： Simple Summary: Caprine arthritis encephalitis is an infectious disease caused by the caprine arthritis encephalitis virus (CAEV). This virus can impair the quality of goat's milk, lamb, and other goat products. Most goats are latently infected with this virus and become long-term carriers, so the development of a rapid and accurate diagnostic method is necessary. Therefore, we constructed a TaqMan real-time quantitative polymerase chain reaction (RT-qPCR) assay and used it to detect CAEV in sheep in eastern China. We found that the prevalence of CAEV infection in sheep in eastern China was 0.77%. This method yields positive results when testing positive references and negative results when testing negative controls or other viruses. The consistency of these results was confirmed through repeating the experiment multiple times. It demonstrated excellent sensitivity, specificity, and reproducibility and holds great promise for its potential application in clinical and field samples in the future. 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-2) was >0.999, and the sensitivity of the assay to the plasmid-carried partial gag gene was approximately 10 copies/mu L, 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.

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