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大豆科学  2007 

进境大豆种子中烟草环斑病毒的快速检测

DOI: 10.3969/j.issn.1000-9841.2007.05.023, PP. 748-751

Keywords: 大豆,烟草环斑病毒,快速检测,RT-PCR,半巢式PCR

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Abstract:

烟草环斑病毒(Tobaccoringspotvirus,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359bp特异性产物。该产物经半巢式PCR进一步验证得到206bp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。

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