title: Development of a SYBR Green I real-time PCR for the detection of the orf virus.
abstract: 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.
authors.affiliation: College of Animal Science and Technology, Anhui Agricultural University, Hefei, 230036, People's Republic of China. wangyong119@ahau.edu.cn.
authors.forename: Yong
authors.initials: Y
authors.lastname: Wang
authors.affiliation: College of Animal Science and Technology, Anhui Agricultural University, Hefei, 230036, People's Republic of China.
authors.forename: Kankan
authors.initials: K
authors.lastname: Yang
authors.affiliation: College of Animal Science and Technology, Anhui Agricultural University, Hefei, 230036, People's Republic of China.
authors.forename: Caixia
authors.initials: C
authors.lastname: Bai
authors.affiliation: College of Animal Science and Technology, Anhui Agricultural University, Hefei, 230036, People's Republic of China.
authors.forename: Dongdong
authors.initials: D
authors.lastname: Yin
authors.affiliation: 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.
authors.forename: Gang
authors.initials: G
authors.lastname: Li
authors.affiliation: College of Animal Science and Technology, Anhui Agricultural University, Hefei, 230036, People's Republic of China.
authors.forename: Kezong
authors.initials: K
authors.lastname: Qi
authors.affiliation: College of Animal Science and Technology, Anhui Agricultural University, Hefei, 230036, People's Republic of China.
authors.forename: Guijun
authors.initials: G
authors.lastname: Wang
authors.affiliation: Municipal Key Laboratory of Virology, Ningbo Municipal Center for Disease Control and Prevention, Ningbo, 315010, People's Republic of China. liyd@nbcdc.org.cn.
authors.forename: Yongdong
authors.initials: Y
authors.lastname: Li
keywords: Detection; Orf virus; Real-time PCR; SYBR Green I