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浙江大学学报(农业与生命科学版)  2010, Vol. 36 Issue (5): 491-496    DOI: 10.3785/j.issn.1008-9209.2010.05.003
生物科学与技术     
一种豇豆重花叶病毒IC-RT real-time PCR检测方法的建立
李彬,粟寒,吴翠萍,周明华,安榆林;
江苏出入境检验检疫局,江苏南京210001;
Development of IC- RT real- time PCR method for detection of Cowpea severe mosaic virus
LI Bin,SU Han,WU Cui- ping,ZHOU Ming-hua,AN Yu-lin
Jiangsu Entry-Exit Inspection and Quarantine Bureau,Nanjing210001,China
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摘要: 为建立豇豆重花叶病毒(Cowpea severe mosaic virus,CPSMV)免疫捕获-反转录-实时荧光(IC-RT real-time)PCR分子检测方法,利用从 ATCC和 DSMZ引进的 CPSMV标准毒株,先建立 CPSMV RT real-time PCR检测方法 ;在此基础上,建立用于 CPSMV检测的IC-RT real-time PCR方法 。结果表明 :IC-RT real-time PCR方法的灵敏度可达500 pg叶组织 ;该方法具有特异性好、灵敏度高、操作简便 等优点,省去了 RNA的提取过程,适用于豇豆等豆类种子及种苗中对豇豆重花叶病毒的快速检测 。
Abstract: A method of immuno-capture reverse transcript real-time(IC-RT real-time) PCR assay was developed for detection of Cowpea severe mosaic virus (CPSMV). By introducing standard CPSMV strains from ATCC and DSMZ as materials,RT real-time PCR method was constituted firstly,which showed good amplification reaction and specificity . Based on the RT real-time PCR,IC-RT real-time PCR detection method for CPSMV was set up . The sensitivity of the assay could reach 500 pg leaf tissues . Meanwhile,the IC-RT real-time PCR assay was characteristic of good specificity,high sensitivity,convenient operation,and avoided RNA extraction,which was suitable for rapid detection of CPSMV from legume seeds and seedlings such as cowpea .
出版日期: 2010-09-13
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李彬
粟寒
吴翠萍
周明华
安榆林

引用本文:

李彬,粟寒,吴翠萍,周明华,安榆林;. 一种豇豆重花叶病毒IC-RT real-time PCR检测方法的建立[J]. 浙江大学学报(农业与生命科学版), 2010, 36(5): 491-496.

LI Bin,SU Han,WU Cui- ping,ZHOU Ming-hua,AN Yu-lin. Development of IC- RT real- time PCR method for detection of Cowpea severe mosaic virus. Journal of Zhejiang University: Agric. & Life Sci., 2010, 36(5): 491-496.

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http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2010.05.003        http://www.zjujournals.com/agr/CN/Y2010/V36/I5/491

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