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以进境种球中截获的带毒洋水仙和郁金香为试验材料,建立了ArMV的免疫捕获RT-PCR、巢式PCR和Real-timePCR方法,并比较了几种检测方法的灵敏度。DAS-ELISA的检测灵敏度较低,为1 mg洋水仙或10 mg郁金香带毒种球,而各种PCR方法的灵敏度可高于DAS-ELISA 100倍以上,其中Real-time PCR检测的灵敏度最高,可从20 ng洋水仙或2μg郁金香的带毒种球中检出ArMV。鉴于DAS-ELISA灵敏度较低,建议在用ELISA初筛时,如样品OD405值与阴性对照OD405值之比在2.0左右时需要再用分子方法加以确证,以防漏检。
The ArMV immunocapture RT-PCR, nested PCR and Real-timePCR methods were established with the poisoned daffodils and tulips intercepted in the entry ball, and the sensitivity of several detection methods was compared. The detection sensitivity of DAS-ELISA was 1 mg daffodil or 10 mg tulip tape, while the sensitivity of various PCR methods was more than 100 times higher than that of DAS-ELISA. The detection sensitivity of Real-time PCR was the highest, ArMV can be detected from poisoned bulbs of 20 ng daffodils or 2 μg of tulips. In view of the lower sensitivity of DAS-ELISA, it is recommended that when screening by ELISA, such as the sample OD405 value and the negative control OD405 value ratio of about 2.0 and then need to be confirmed by molecular methods to prevent missed.