【摘要】 采用环介导等温扩增技术(loop-mediated isothermal amplification,LAMP),建立了草莓潜隐环斑病毒(Strawberry latent ringspot virus,SLRSV)检测方法。根据SLRSV外壳蛋白编码基因上的8个位点,共设计了6条引物,通过逆转录环介导等温扩增得到特征性的瀑布状条带。特异性试验表明,引物对SLRSV的检测具有良好的特异性;灵敏度试验显示RT-LAMP的灵敏度比普通RT-PCR高出100倍。该方法无需特殊的试剂和设备,只需在水浴锅中65℃等温扩增,整个检测周期约1.5h,结果采用SYBR green I染色显示,易于观察和判定。更多还原

【Abstract】 A loop-mediated isothermal amplification(LAMP)method was established for detection of Strawberry latent ringspot virus(SLRSV)in the study.Based on eight sites on SLRSV coat protein coding genes,six primers were designed,and characteristic waterfall-like bands were obtained by RT-LAMP.Specificity tests showed that the primers are specific for SLRSV.The sensitivity of RT-LAMP is 100-fold higher than that of RT-PCR.The method does not need special equipment and reagents and the reaction condition is that the incubation temperature is 65℃ and the reaction time 1.5 h.The results can be easily observed by using SYBR green I.更多还原