Development and Application of a Real-time Fluorescence Quantitative PCR for Detection of Orf Virus

LIN Yu-sheng; JIANG Jin-xiu; ZHANG Jing-peng; JIANG Bin; YOU Wei; HU Qi-lin

doi:10.19303/j.issn.1008-0384.2018.04.002

Volume 33 Issue 4

Mar. 2019

Turn off MathJax

Article Contents

Article Navigation > Fujian Journal of Agricultural Sciences > 2018 > 33(4): 341-345

LIN Yu-sheng, JIANG Jin-xiu, ZHANG Jing-peng, JIANG Bin, YOU Wei, HU Qi-lin. Development and Application of a Real-time Fluorescence Quantitative PCR for Detection of Orf Virus[J]. Fujian Journal of Agricultural Sciences, 2018, 33(4): 341-345. doi: 10.19303/j.issn.1008-0384.2018.04.002

Citation:

LIN Yu-sheng, JIANG Jin-xiu, ZHANG Jing-peng, JIANG Bin, YOU Wei, HU Qi-lin. Development and Application of a Real-time Fluorescence Quantitative PCR for Detection of Orf Virus[J]. Fujian Journal of Agricultural Sciences, 2018, 33(4): 341-345. doi: 10.19303/j.issn.1008-0384.2018.04.002

Citation:

LIN Yu-sheng, JIANG Jin-xiu, ZHANG Jing-peng, JIANG Bin, YOU Wei, HU Qi-lin. Development and Application of a Real-time Fluorescence Quantitative PCR for Detection of Orf Virus[J]. Fujian Journal of Agricultural Sciences, 2018, 33(4): 341-345. doi: 10.19303/j.issn.1008-0384.2018.04.002

PDF( 1529 KB)

Development and Application of a Real-time Fluorescence Quantitative PCR for Detection of Orf Virus

doi: 10.19303/j.issn.1008-0384.2018.04.002

Institute of Animal Husbandry & Veterinary Medicine, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian 350013, China

Received Date: 2018-01-25
Rev Recd Date: 2018-03-11
Publish Date: 2018-04-01

Abstract

Abstract

The study established a SYBR Green I real-time fluorescence quantitative PCR assay for detecting orf virus. Using Beacon Designer 7.9 software to design a pair of specific primers based on the VIR gene of the orf virus in the Genbank (accession number.KF666563.1). The result showed that it would specifically detect orf virus, but wouldn't detect other sheep and goat bacteria and viruses. The detection limit of the method was 100 copies·μL^-1 and the detected Ct values of intra-and inter-variation were all less than 2%. Using the assay to detect 39 clinical samples, the positive rate was 94.9% (37/39). The above results indicated that the assay had a high specificity, sensitivity and repeatability, which could be used to detect orf virus and conduct epidemiological investigation.
- orf virus,
- SYBR Green I,
- real-time fluorescence quantitative PCR

FullText(HTML)

References(17)

References

[1]	杨海波, 孟庆玲, 乔军, 等.羊口疮病毒新疆流行株的分离鉴定及其遗传进化分析[J].西北农林大学学报(自然科学版), 2015, 43(2):14-22. http://www.cnki.com.cn/Article/CJFDTotal-XBNY201502004.htm
[2]	余波, 冉懋韬, 谭诗文, 等.羊传染性脓疱病毒PCR检测方法的建立及CBP基因的序列分析[J].畜牧与兽医, 2013, 45(7):30-34. http://www.cqvip.com/QK/95129X/201307/46504644.html
[3]	JIDA L, DEGUANG S, WENQI H, et al. Rapid detection of orf virus by loop-mediated isothermal amplification based on the DNA polymerase gene[J]. Arch Virol, 2013, 158:793-798. doi: 10.1007/s00705-012-1526-1
[4]	GALLINA I, Dal POZZO F, Mc INNES C, et al. A real time PCR assay for the detection and quantification of orf virus[J]. J Virol Meth, 2006, 134(1):140-145. http://cn.bing.com/academic/profile?id=1559bab1272828ac93b88bfe7f6345e7&encoded=0&v=paper_preview&mkt=zh-cn
[5]	GUANGXIANG W, YOUJUN S, YANHUA W, et al. Comparison of a loop-mediated isothermal amplification for orf virus with quantitative real-time PCR[J]. Virol J, 2013(10):138-147. http://cn.bing.com/academic/profile?id=bd414a97b16ef6523d0235526cb55d9f&encoded=0&v=paper_preview&mkt=zh-cn
[6]	HAIG D M, MCINNES C J, THOMSOn J, et al. The orf virus OV20.0L gene product is involved in interferon resistance and inhibits an interferon-inducible, double-stranded RNA-dependent kinase[J]. Immunology, 1988, 93(3):335-340. http://cn.bing.com/academic/profile?id=ccbc6fad28b60cfc8e0a06ae294950c0&encoded=0&v=paper_preview&mkt=zh-cn
[7]	KOTTARIDI C, NOMIKOU K, TEODORI L, et al. Phylogenetic correlation of Greek and Italian orf virus isolates based on VIR gene[J]. Vet Microbiol, 2006, 116(4):310-316. doi: 10.1016/j.vetmic.2006.04.020
[8]	GUO J, ZHANG Z, EDWARDS J F, et al. Characterization of a North American orf virus isolated from a goat with persistent proliferative dermatitis[J]. Vir Res, 2003, 93(2):169-179. doi: 10.1016/S0168-1702(03)00095-9
[9]	CHAN K W, YANG C H, LIN J W, et al. Phylogenetic analysis of parapoxviruses and the C-terminal heterogeneity of viral ATPase proteins[J].Gene, 2009, 432(1-2):44-53. doi: 10.1016/j.gene.2008.10.029
[10]	林裕胜, 江锦秀, 张靖鹏, 等.丝状支原体山羊亚种SYBR Green I qRT-PCR快速检测方法的建立[J].农业生物技术学报, 2017, 25(11):1895-1902. http://www.cqvip.com/QK/93164X/201607/669461466.html
[11]	颜新敏, 储岳峰, 吴国华, 等.山羊痘、羊口疮及山羊传染性胸膜肺炎混合感染的检测与分析[J].中国兽医学报, 2011, 31(6):809-813. http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=zgsyxb201106009
[12]	林裕胜, 江锦秀, 江斌, 等.福建省羊传染性脓疱病毒F1L、B2L和VIR基因的遗传变异分析[J], 动物医学进展, 2017, 38(2):1-6. http://cpfd.cnki.com.cn/Article/CPFDTOTAL-ZGXJ200805001087.htm
[13]	LI H, ZHU X, ZHENG Y, et al. Phylogenetic analysis of two Chinese orf virus isolates based on sequences of B2L and VIR genes[J]. Arch Virol, 2013, 158(7):1477-1485. doi: 10.1007/s00705-013-1641-7
[14]	OEM J K, ROH I S, LEE K H, et al. Phylogenetic analysis and characterization of Korean orf virus from dairy goats:case report[J]. Virol J, 2009(6):167. http://cn.bing.com/academic/profile?id=d2dbb3ed505ca939ad6f6d69a9d782fb&encoded=0&v=paper_preview&mkt=zh-cn
[15]	鲜思美, 张素辉, 杨钰, 等.羊口疮病毒SYBR Green I实时荧光定量PCR的建立及应用[J].贵州农业科学, 2014, 42(6):104-108.
[16]	姚俊, 李富祥, 彭海生, 等.羊口疮病毒TaqMan实时荧光定量PCR检测方法的建立及应用[J].动物医学进展, 2012, 33(11):31-36. doi: 10.3969/j.issn.1007-5038.2012.11.007
[17]	GALLINA L, DALPOZZO F, MCINNES C J, et al. A real time PCR assay for the detection and quantification of orfvirus[J]. J Virol Methods, 2006, 134(1-2):140-145. http://cn.bing.com/academic/profile?id=1559bab1272828ac93b88bfe7f6345e7&encoded=0&v=paper_preview&mkt=zh-cn