| 恒温隔绝式荧光PCR检测非洲猪瘟病毒核酸方法的建立 |
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| 引用本文： | 陈雪蓉,徐林,王远微,汤承,岳华.恒温隔绝式荧光PCR检测非洲猪瘟病毒核酸方法的建立[J].中国动物检疫,2022,39(3):80-84. |
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| 作者姓名： | 陈雪蓉 徐林 王远微 汤承 岳华 |
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| 作者单位： | 西南民族大学畜牧兽医学院;甘孜州动物疫病预防控制中心 |
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| 基金项目： | 四川省科技计划重点研发项目（2018NZ0151-3）；国家农业产业技术体系四川兽药创新团队项目（SCCXTD-2020-18）；西南民族大学动物医学实验室平台建设项目（2020PTJS29） |
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| 摘 要： | 恒温隔绝式荧光PCR(iiPCR)是一种简便、快速及可使用于现场检测的荧光PCR方法.本研究采用农业农村部推荐的检测非洲猪瘟病毒(African swine fever virus,ASFV)核酸的荧光定量PCR方法的引物和探针,通过对引物、探针、Taq酶等反应体系的优化,建立了基于恒温隔绝式荧光PCR的ASFV检测方...
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| 关 键 词： | 非洲猪瘟病毒 恒温隔绝式荧光PCR 现场检测 |
Development of an Insulated Isothermal PCR Assay for Detecting African Swine Fever Virus |
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| Chen Xuerong,Xu Lin,Wang Yuanwei,Tang Cheng,Yue Hua.Development of an Insulated Isothermal PCR Assay for Detecting African Swine Fever Virus[J].China Journal Of Animal Quarantine,2022,39(3):80-84. |
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| Authors: | Chen Xuerong Xu Lin Wang Yuanwei Tang Cheng Yue Hua |
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| Institution: | (College of Animal and Veterinary Sciences,Southwest Minzu University,Chengdu,Sichuan 610041,China;Ganzi Center for Animal Disease Prevention and Control,Kangding,Sichuan 626000,China) |
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| Abstract: | Insulated isothermal PCR(iiPCR)is a simple and rapid fluorescent PCR assay to be used for field detection.In the study,the primers and probes of the fluorescence quantitative PCR for detecting African swine fever virus(ASFV)recommended by the Ministry of Agricultural and Rural Affairs were used,after optimization of reaction system including primers,probes and Taq enzyme,an iiPCR was established.By the established method,ASFV nucleic acids could be specifically detected,and negative results were obtained for other six viruses that usually infected pigs;the detection limit was 2.55 copies/μL,and the variable coefficients within and between groups were less than 2%.For 100 clinical samples,the test results by the established method were consistent with those by fluorescence quantitative PCR recommended by the Ministry of Agricultural and Rural Affairs.In conclusion,the specificity,stability and sensitivity of the established method were consistent with those of the traditional fluorescence quantitative PCR,which realized the standardization and field detection of ASFV,and provided a reliable tool for ASFV rapid detection. |
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| Keywords: | ASFV ⅱPCR field detection |
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