用于鉴别检测山羊痘病毒和绵羊痘病毒的双重PCR技术的开发
Development of duplex PCR for differential detection of goatpox and sheeppox viruses.
作者信息
Zhao Zhixun, Wu Guohua, Yan Xinmin, Zhu Xueliang, Li Jian, Zhu Haixia, Zhang Zhidong, Zhang Qiang
机构信息
Key Laboratory of Animal virology of the Ministry of Agriculture, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, CAAS, Lanzhou, People's Republic of China.
出版信息
BMC Vet Res. 2017 Aug 31;13(1):278. doi: 10.1186/s12917-017-1179-0.
BACKGROUND
Clinically, sheeppox and goatpox have the same symptoms and cannot be distinguished serologically. A cheaper and easy method for differential diagnosis is important in control of this disease in endemic region.
METHODS
A duplex PCR assay was developed for the specific differential detection of Goatpox virus (GTPV) and Sheeppox virus (SPPV), using two sets of primers based on viral E10R gene and RPO132 gene.
RESULTS
Nucleic acid electrophoresis results showed that SPPV-positive samples appear two bands, and GTPV-positive samples only one stripe. There were no cross-reactions with nucleic acids extracted from other pathogens including foot-and-mouth disease virus, Orf virus. The duplex PCR assay developed can specially detect SPPV or GTPV present in samples (n = 135) collected from suspected cases of Capripox.
CONCLUSIONS
The duplex PCR assay developed is a specific and sensitive method for the differential diagnosis of GTPV and SPPV infection, with the potential to be standardized as a detection method for Capripox in endemic areas.
背景
临床上，绵羊痘和山羊痘具有相同症状，且无法通过血清学进行区分。在流行地区，一种更廉价且简便的鉴别诊断方法对于控制这种疾病至关重要。
方法
基于病毒E10R基因和RPO132基因设计两组引物，建立了一种用于特异性鉴别检测山羊痘病毒（GTPV）和绵羊痘病毒（SPPV）的双重PCR检测方法。
结果
核酸电泳结果显示，SPPV阳性样本出现两条带，而GTPV阳性样本仅出现一条带。与从包括口蹄疫病毒、羊口疮病毒等其他病原体中提取的核酸无交叉反应。所建立的双重PCR检测方法能够特异性检测从疑似羊痘病例采集的样本（n = 135）中存在的SPPV或GTPV。
结论
所建立的双重PCR检测方法是一种用于鉴别诊断GTPV和SPPV感染的特异性和灵敏性方法，有潜力被标准化为流行地区羊痘的检测方法。
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