番茄斑萎病毒RT-RPA检测方法的建立

Development of RT-RPA for detection of tomato spotted wilt virus

为了快速、准确地鉴定番茄斑萎病毒,根据GenBank中韩国的番茄斑萎病毒株系(序列号:KC261961)ssRNA-S基因组编码保守外壳蛋白基因序列设计特异性引物TSWV 5F/5R,建立了番茄斑萎病毒RT-RPA检测方法。特异性检测结果表明,仅番茄斑萎病毒样品扩增出215 bp目标条带,而其他4种病毒样品无扩增出目标条带。灵敏度检测结果表明番茄斑萎病毒RPA检测方法可以达到10-5 cDNA稀释度。

For rapid and accurate detection of tomato spotted wilt virus(TSWV),the primer TSWV 5F/5R was designed based on ssRNA-S sequence of Korean TSWV strain KC261961 in the GenBank to established recombinase polymerase amplification.Specificity determination showed that only TSWV sample amplified a 215 bp target band,while other samples didn’t amplify target band.Sensitity determination results demonstrated that RPA method could reach tomato spotted wilt virus cDNA dilution to 10-5.