| A型猪塞内卡病毒TaqMan荧光定量PCR方法的建立 |
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| 引用本文： | 李秀博,刘存,鄢明华,崔玉东,崔尚金.A型猪塞内卡病毒TaqMan荧光定量PCR方法的建立[J].中国动物传染病学报,2019(3):33-38. |
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| 作者姓名： | 李秀博 刘存 鄢明华 崔玉东 崔尚金 |
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| 作者单位： | 黑龙江八一农垦大学动物科技学院;中国农业科学院北京畜牧兽医研究所;农业部兽用药物与诊断技术北京科学观测实验站;天津畜牧兽医研究所 |
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| 基金项目： | 中国农业科学院创新工程项目(ASTIP-IAS15);国家重点研发项目(2017YFD0502300);天津市生猪产业创新技术体系项目(ITTPRS2017003) |
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| 摘 要： | 本研究针对A型猪塞内卡病毒(Senecavirus A, SVA)基因组保守区域设计了特异性引物及探针,建立了快速检测SVA的Taq Man荧光定量PCR方法。结果显示,以构建的重组质粒为标准品建立的Taq Man荧光定量PCR方法,标准曲线具有良好的线性关系,线性相关系数达0.9974;特异性良好,与口蹄疫病毒、猪流行性腹泻病毒、猪传染性胃肠炎病毒、猪瘟病毒、猪伪狂犬病毒不存在交叉反应;对SVA核酸最低检测下限为3.75 copies/μL,而普通PCR最低检测下限为3.75×103 copies/μL;批内和批间的变异系数为均小于5%,重复性良好。本研究建立的SVA TaqMan荧光定量PCR方法为检测猪水疱性疾病病原提供了一种快速、灵敏的检测方法,为开展SVA流行病学调查提供了技术支持。
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| 关 键 词： | A型猪塞内卡病毒 TaqMan荧光定量PCR 诊断 |
DEVELOPMENT AND APPLICATION OF A TAQMAN REAL-TIME PCR ASSAY FOR DETECTION OF PORCINE SENECAVIRUS A |
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| LI Xiu-bo,LIU Cun,YAN Ming-hua,CUI Yu-dong,CUI Shang-jin.DEVELOPMENT AND APPLICATION OF A TAQMAN REAL-TIME PCR ASSAY FOR DETECTION OF PORCINE SENECAVIRUS A[J].Chinese Journal of Animal Infectious Diseases,2019(3):33-38. |
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| Authors: | LI Xiu-bo LIU Cun YAN Ming-hua CUI Yu-dong CUI Shang-jin |
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| Institution: | (College of Animal Science and Technology, Heilongjiang Bayi Agricultural University, Daqing 163319, China;Institute of Animal Science Chinese Academy of Agricultural Sciences, Beijing100193, China;Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Technology of Beijing, Ministry of Agriculture, Beijing 100193, China;Tianjin Institute of Animal Husbandry and Veterinary Science, Tianjin300384, China) |
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| Abstract: | To develop a rapidly diagnostic method for detection of porcine Senecavirus A(SVA), a fluorogenic TaqMan real-time PCR assay was developed in the present study. There was no cross-reaction with foot and mouth disease virus, porcine epidemic diarrhea virus, swine fever virus and pseudorabies virus. The assay was able to detect as low as 3.75 copies and a dynamic range of at least eight orders of magnitude. The results showed that this assay was more sensitive than traditional PCR assay that had a sensitivity of 3.75×103 copies. In addition, this assay was reproducibility with less than 5% of the coefficient of variation within a batch and among batches. In conclusion, the fluorogenic TaqMan real-time PCR assay performed as a fast and sensitive diagnostic tool for detection of SVA that might be used for epidemiological investigation. |
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| Keywords: | Senecavirus A TaqMan real-time PCR diagnosis method |
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