利用斑点杂交法和RT-PCR技术检测甘蔗花叶病毒

Detection of -Sugarcane mosaic virus- by dot hybridization and RT PCR technique

利用地高辛标记不同长度的 DNA探针通过斑点杂交法对甘蔗花叶病毒进行检测 .结果表明 ,可检测出2 0 0 mg稀释度为 1/10 4 病叶中的病毒 .但不同长度的探针表现出一定的差异 ,较长的探针显示出较高的灵敏度 .根据克隆所得的病毒基因序列设计特异引物 ,利用反转录 -聚合酶链式反应的方法进行检测 ,同样得到较好的检测效果 ,但灵敏度略低于杂交检测 .

Different lengthes of digoxigenin labeled DNA probes were used to detect Sugarcane mosaic virus by dot hybridization, and the virus in 200 mg of diseased leaves at the dilution of 1/10 4 was detected, the long probe, however, performed high sensitivity, which revealed the distinguishment between different lengthes of probes. Meanwhile, when the specific primers were designed based on the sequence of the virus gene obtained by cloning and reverse transcription polymerase chain reaction (RT PCR) was used to detect the virus, an ideal result was also acquired but its sensitivity was lower than that of dot hybridization.