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用PBS和种子研磨后的普通病原提取液稀释番茄溃疡病菌纯菌液,并以梯度纯菌液和带菌种子提取液作为模板进行直接PCR和免疫捕捉PCR,比较2种方法在纯菌液和带菌种子提取液中的检测灵敏度,找到一种高特异性、高灵敏度、简便快捷的方法用于检测番茄种子携带的番茄溃疡病菌。结果显示在纯菌液中直接PCR灵敏度为104cfu/mL,免疫捕捉PCR为102cfu/mL;在带菌种子提取液中直接PCR灵敏度为106cfu/mL,免疫捕捉PCR为104cfu/mL;免疫捕捉PCR比直接PCR灵敏度高100倍,尤其在实际种子检测中优势更明显,而且检出时间短,重复性好。
The pure pathogenic bacteria of tomato canker were diluted with ordinary pathogen extracts after grinding with PBS and seeds, and the direct and gradient PCR and immunofluorescent PCR were carried out using the gradient pure bacterial and the extracted seed extract as templates. Seed extract detection sensitivity, to find a high specificity, high sensitivity, simple and quick method for the detection of tomato seeds carried by tomato canker. The results showed that the sensitivity of direct PCR was 104cfu / mL in pure bacterial liquid and 102cfu / mL in immune-capture PCR. The direct PCR sensitivity was 106cfu / mL and the immune capture PCR was 104cfu / mL. PCR sensitivity of 100 times, especially in the actual seed detection advantages are more obvious, and the detection time is short, good repeatability.