CAJ | 학술논문

为了快速有效地检测出植物样品中的番茄黑环病毒(tomato black ring virus, TBRV),根据TBRV 的保守序列设计 RT - LAMP 反应的特异引物,通过试验成功建立了 TBRV 的 RT - LAMP 特异性检测方法。该方法检测快速,约27 min 即可检测到扩增曲线；特异性强,可有效区分同样侵染马铃薯的马铃薯 A 病毒(PVA)、马铃薯 Y 病毒(PVY)、番茄斑萎病毒( TSWV)及番茄环斑病毒(ToRSV)；灵敏度高,比普通 RT - PCR 灵敏度至少高10倍；并可通过肉眼观察反应颜色的变化直接进行结果判断。该方法适用于 TBRV 的快速检测。
위료쾌속유효지검측출식물양품중적번가흑배병독(tomato black ring virus, TBRV),근거TBRV 적보수서렬설계 RT - LAMP 반응적특이인물,통과시험성공건립료 TBRV 적 RT - LAMP 특이성검측방법。해방법검측쾌속,약27 min 즉가검측도확증곡선；특이성강,가유효구분동양침염마령서적마령서 A 병독(PVA)、마령서 Y 병독(PVY)、번가반위병독( TSWV)급번가배반병독(ToRSV)；령민도고,비보통 RT - PCR 령민도지소고10배；병가통과육안관찰반응안색적변화직접진행결과판단。해방법괄용우 TBRV 적쾌속검측。
A one-step reverse transcription loop-mediated isothermal amplification( RT-LAMP) method for detection of tomato black ring virus(TBRV) was developed,providing a quick and effective way to de-tect TBRV in plant materials. Using the Primer Explorer software,RT-LAMP primers were designed based on a conserved region of TBRV. The method only needed about twenty-seven minutes for obtaining of the amplification curve. The amplification curve result of specificity test using TBRV,potato virus A(PVA), potato virus Y(PVY),tomato spotted wilt virus(TSWV),tomato ringspot virus(ToRSV),healthy potato leaves and water as template showed that the assay could amplify TBRV specifically,in consistent with the fluorescent detection reagent(FDR) reaction result. The sensitivity comparison showed that the RT-LAMP method was at least 10 times more sensitive than conventional reverse transcriptase polymerase chain reac-tion(RT-PCR) using serial dilutions of total RNA extracts. In addition,the result could be judged direct-ly by observing the changes in color of the reaction using the RT-LAMP method. Overall,RT-LAMP is a sensitive,simple,specific diagnostic tool,which should be of value in more accurate and quick determina-tion of the distribution of TBRV.