[1]闻伟刚,崔俊霞,盛蕾.进境大豆种子中烟草环斑病毒的快速检测[J].大豆科学,2007,26(05):748-751.[doi:10.3969/j.issn.1000-9841.2007.05.023]
　WEN Wei-gang,CUI Jun-xia,SHENG Lei.RAPID DETECTION OF TOBACCO RINGSPOT VIRUS IN IMPORTED SOYBEAN SEEDS[J].Soybean Science,2007,26(05):748-751.[doi:10.3969/j.issn.1000-9841.2007.05.023]
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进境大豆种子中烟草环斑病毒的快速检测

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第26卷 期数: 2007年05期 页码: 748-751 栏目: 出版日期: 2007-10-25
Title:
RAPID DETECTION OF TOBACCO RINGSPOT VIRUS IN IMPORTED SOYBEAN SEEDS
文章编号:
1000-9841(2007)05-00748-04
作者:
闻伟刚; 崔俊霞; 盛蕾
宁波出入境检验检疫局，宁波 315012
Author(s):
WEN Wei-gang; CUI Jun-xia; SHENG Lei
Ningbo Entry-exit Inspection and Quarantine Bureau，Ninbo 315012
关键词:
大豆; 烟草环斑病毒; 快速检测; RT-PCR; 半巢式PCR
Keywords:
Soybean; Tobacco ringspot virus; Rapid detection ; RT-PCR; Semi-nested PCR
分类号:
S41-31
DOI:
10.3969/j.issn.1000-9841.2007.05.023
文献标志码:
A
摘要:
烟草环斑病毒（Tobacco ringspot virus，TRSV）是进境大豆种子必须检疫的项目，属于我国禁止入境的二类检疫性有害生物，本文建立了大豆种子中TRSV的快速检测方法。采用Trizol试剂直接提取大豆种子总RNA，通过RT-PCR扩增得到359 bp特异性产物。该产物经半巢式PCR进一步验证得到206 bp特异性片段，经测序及同源性比较表明，片段序列与GenBank中登录的TRSV外壳蛋白基因部分序列存在92％～96％的同源性，而未见与其它基因序列存在同源性。同时，大豆种子样品经DAS-ELISA复核检测结果显示TRSV阳性反应。因此，判定该批进境大豆种子携带TRSV。
Abstract:
Tobacco ringspot virus(TRSV) is the second level quarantine pest which should be tested in imported soybean.In this paper，a rapid detection method was established which used Trizol for extracting total RNA of soybean seeds directly and adopted RT-PCR for amplification，a 359 bp special band was obtained.Through further verification of semi-nested PCR and sequence homologous comparation，a 206 bp special product was obtained and had 92%~96% identities with TRSV sequence of coat protein genes published in GeneBank.The homology to other genetic sequences of plant viruses were not found.Meanwhile，the soybean seeds sample showed TRSV positive reaction when checked by DAS-ELISA.Thus，the soybean seeds sample was determined to contain TRSV.

参考文献/References:

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备注/Memo

基金项目：宁波出入境检验检疫局科技项目（甬K34-2006）

作者简介：闻伟刚（1975－），男，博士研究生，主要从事疫病分子生物学检测工作。

更新日期/Last Update: 2014-10-20