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浙江大学学报（农业与生命科学版）
Journal of Zhejiang University: Agric. & Life Sci.  2010, Vol. 36 Issue (5): 491-496    DOI: 10.3785/j.issn.1008-9209.2010.05.003
Biological sciences & biotechnology     
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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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 .

Published: 13 September 2010
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LI Bin
SU Han
WU Cui- ping
ZHOU Ming-hua
AN Yu-lin
Cite this article:

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


一种豇豆重花叶病毒IC-RT real-time PCR检测方法的建立

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 。
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