柑橘叶斑驳病毒的RT-LAMP检测
Acta Horticulturae Sinica(2020)
Abstract
建立了柑橘叶斑驳病毒(citrus leaf blotch virus, CLBV)的RT-LAMP (Reverse transcription loop-mediated isothermal amplification)检测方法,并对其灵敏度和准确性进行验证.根据CLBV的外壳蛋白序列设计引物,筛选出特异性较好的一组引物,进行扩增时间及温度等条件优化.优化后反应体系可在65 ℃条件下,60 min内完成扩增.所建立体系特异性好,灵敏度高,最低可从23.2 pg · μL-1的模板中检测出CLBV,扩增结果可通过加入SYBR green I染料方法直接观察.用RT-LAMP方法对田间采集的120份柑橘样品进行检测,不同来源样品检出率在0~15%之间,与普通RT-PCR方法检出结果一致.
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