Wang Yong, Yang Kankan, Bai Caixia, Yin Dongdong, Li Gang, Qi Kezong, Wang Guijun, Li Yongdong
College of Animal Science and Technology, Anhui Agricultural University, Hefei, 230036, People's Republic of China.
State Key Laboratory of Animal Nutrition, Beijing Institute of Animal Science and Veterinary Medicine, Chinese Academy of Agricultural Sciences, Beijing, 100193, People's Republic of China.
AMB Express. 2017 Dec;7(1):21. doi: 10.1186/s13568-016-0322-9. Epub 2017 Jan 7.
Orf is a non-systemic, ubiquitous disease of sheep and goats caused by the orf virus (ORFV). ORFV occasionally causes cutaneous lesions in humans in contact with infected animals. In the present study, a real-time PCR method was established for detection of ORFV using the fluorescent chimeric dye SYBR Green I. Specific primers were designed to target a highly conserved region of the ORFV B2L gene. This method was able to detect a minimum of 20 copies of ORFV genomic DNA. The results showed no cross-reactions with other common DNA viruses. The time required for the test was approximately 1.5 h. Clinical test samples showed that this method was faster and had a higher sensitivity than traditional PCR. In conclusion, this novel, real-time PCR-based assay provides a rapid, sensitive, and specific method for ORFV detection. This test provides improved technical support for studies regarding the clinical diagnosis and epidemiology of ORFV.
羊口疮是由羊口疮病毒（ORFV）引起的一种非全身性、广泛存在于绵羊和山羊中的疾病。ORFV偶尔会在与受感染动物接触的人类身上引起皮肤损伤。在本研究中，建立了一种使用荧光嵌合染料SYBR Green I检测ORFV的实时PCR方法。设计了特异性引物靶向ORFV B2L基因的高度保守区域。该方法能够检测到至少20拷贝数的ORFV基因组DNA。结果显示与其他常见DNA病毒无交叉反应。该检测所需时间约为1.5小时。临床试验样本表明，该方法比传统PCR更快且灵敏度更高。总之，这种基于实时PCR的新检测方法为ORFV检测提供了一种快速、灵敏且特异的方法。该检测为羊口疮病毒临床诊断和流行病学研究提供了更好的技术支持。
