逆转录环介导等温扩增技术检测草莓潜隐环斑病毒的研究  

Detection of Strawberrylatentringspotvirusby RT-LAMP

作　　者：张吉红[1] 余澍琼[1] 徐瑛[1] 张慧丽[1] 陈先锋[1] 陈剑平[2] 陈炯[3] 

机构地区：[1]宁波出入境检验检疫局,宁波315012 [2]浙江省农业科学院,杭州310021 [3]宁波大学,宁波315211

出　　处：《植物保护》2013年第6期74-77,共4页Plant Protection

基　　金：宁波市科研项目(2011C50080);国家质检总局项目(2013IK279);质检公益性行业科研专项(201010256);浙江省重点科技创新团队(2010R50028)

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。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.

关 键 词：草莓潜隐环斑病毒 环介导等温扩增 检测 

分 类 号：S436.639[农业科学—农业昆虫与害虫防治]