| 双重胶体金层析法快速检测番茄环斑病毒和烟草环斑病毒RT-PCR扩增子 |
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| 引用本文： | 曹成,魏梅生,张永江.双重胶体金层析法快速检测番茄环斑病毒和烟草环斑病毒RT-PCR扩增子[J].中国农学通报,2011,27(22):197-201. |
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| 作者姓名： | 曹成 魏梅生 张永江 |
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| 作者单位： | 1. 河南工业大学生物工程学院,郑州,450001
2. 中国检验检疫科学研究院动植物检疫研究所,北京,100029 |
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| 基金项目： | 国家质量监督检验检疫总局课题“检疫性植物病毒核酸层析检测技术研究”(2009IK322) |
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| 摘 要： | 番茄环斑病毒和烟草环斑病毒是我国进境检疫性有害生物，本研究建立了单个和双重胶体金层析快速检测方法，在15min即可获得对双标记PCR产物的检测结果。单个胶体金层析是在一张层析膜上点上荧光素(或地高辛)的单克隆抗体作为T检测点，经生物素－荧光素（或生物素－地高辛）双标记的RT-PCR扩增产物可在T点上被检测到。双重胶体金层析是在一张层析膜上分别点上荧光素和地高辛的单克隆抗体作为T1和T2检测点，经生物素－荧光素和生物素－地高辛双标记的两种病毒的RT-PCR扩增产物可分别在T1和T2点上被检测到。
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| 关 键 词： | 氮肥利用率 氮肥利用率 15N示踪法 综述 |
| 收稿时间： | 2011/3/15 0:00:00 |
| 修稿时间： | 2011/3/24 0:00:00 |
Duplex Detection of Tomato Ringspot Virus and Tobacco Ringspot Virus RT-PCR Amplicons by Dipstick Assay |
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| Cao Cheng,Wei Meisheng,Zhang Yongjiang,Li Guifen,Wu Xingquan.Duplex Detection of Tomato Ringspot Virus and Tobacco Ringspot Virus RT-PCR Amplicons by Dipstick Assay[J].Chinese Agricultural Science Bulletin,2011,27(22):197-201. |
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| Authors: | Cao Cheng Wei Meisheng Zhang Yongjiang Li Guifen Wu Xingquan |
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| Institution: | Cao Cheng 1,Wei Meisheng 2,Zhang Yongjiang 2,Li Guifen 2,Wu Xingquan 1(1 College of Bioengineering,Henan University of Technology,Zhengzhou 450001,2 Institute of Animal and Plant Quarantine,Chinese Academy of Inspection and Quarantine,Beijing 100029) |
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| Abstract: | Both Tomato ringspot virus(ToRSV)and Tobacco ringspot virus(TRSV) are quarantine pests for China.We have developed dipstick assay for detection of RT-PCR amplicons of the two viruses within 15 minutes.The single dipstick assay is performed by labeling primers of the virus with biotin and fluorescein(or digoxigenin).Monoclonal antibody against fluorescein(or digoxigenin) is spotted on nitrocellulose membrane as T dot.The dual-labeled(biotin-fluorescein or biotin-digoxigenin) RT-PCR amplicons can be detected ... |
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| Keywords: | ToRSV TRSV RT-PCR colloidal gold duplex detection |
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