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

被引：26

作者：

Wang, Yong ^{[1
]}

Yang, Kankan ^{[1
]}

Bai, Caixia ^{[1
]}

Yin, Dongdong ^{[1
]}

Li, Gang ^{[3
]}

Qi, Kezong ^{[1
]}

Wang, Guijun ^{[1
]}

Li, Yongdong ^{[2
]}

机构：

[1] Anhui Agr Univ, Coll Anim Sci & Technol, Hefei 230036, Peoples R China

[2] Ningbo Municipal Ctr Dis Control & Prevent, Municipal Key Lab Virol, Ningbo 315010, Peoples R China

[3] Chinese Acad Agr Sci, State Key Lab Anim Nutr, Beijing Inst Anim Sci & Vet Med, Beijing 100193, Peoples R China

来源：

AMB EXPRESS | 2017年 / 7卷

基金：

中国国家自然科学基金;

关键词：

Detection; Orf virus; Real-time PCR; SYBR Green I; POLYMERASE-CHAIN-REACTION; PHYLOGENETIC ANALYSIS; DIAGNOSIS; IDENTIFICATION; AMPLIFICATION; OUTBREAK; GOATS; SHEEP;

D O I：

10.1186/s13568-016-0322-9

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

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.

引用

页数：6

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