Development of a SYBR Green I real-time PCR for the detection of the orf virus

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• 作者：Yong Wang ; Kankan Yang ; Caixia Bai ; Dongdong Yin ; Gang Li ; Kezong Qi…
• 关键词：Detection ; Orf virus ; Real ; time PCR ; SYBR Green I
• 刊名：AMB Express
• 出版年：2017
• 出版时间：December 2017
• 年：2017
• 卷：7
• 期：1
• 全文大小：1506KB
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Microbiology; Microbial Genetics and Genomics; Biotechnology;
• 出版者：Springer Berlin Heidelberg
• ISSN：2191-0855
• 卷排序：7

文摘

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.