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首页> 外文期刊>Plant Pathology >A multiplex RT-PCR for the detection of Potato yellow vein virus, Tobacco rattle virus and Tomato infectious chlorosis virus in potato with a plant internal amplification control
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A multiplex RT-PCR for the detection of Potato yellow vein virus, Tobacco rattle virus and Tomato infectious chlorosis virus in potato with a plant internal amplification control

机译：多重RT-PCR用于植物内部扩增控制，用于检测马铃薯中的马铃薯黄脉病毒，烟草嘎嘎病毒和番茄感染性绿化病毒

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A multiplex reverse transcriptase polymerase chain reaction (RT-PCR) assay was developed using published primers to simultaneously detect three viruses infecting potato (Potato yellow vein virus, Tomato infectious chlorosis virus and Tobacco rattle virus), including an internal amplification control. The characteristics of the published primers were analysed to see if they could be used for multiplex RT-PCR. Amplification of the three target viruses and a plant internal control was then optimized by adding bovine serum albumin during cDNA synthesis, increasing the PCR extension time, reducing the PCR extension temperature and optimizing the concentration of each pair of primers. This optimization produced a multiplex assay with a detection sensitivity of the same order as simplex RT-PCR. This approach provides a simple and convenient way to develop multiplex assays using published primers directly rather than designing new primers. The multiplex RT-PCR is a sensitive and cost-effective method for detecting multiple viruses in potato and can be used for quarantine and certification programmes.
机译：使用公开的引物开发了一种多重逆转录酶聚合酶链反应（RT-PCR）分析方法，可同时检测三种感染马铃薯的病毒（马铃薯黄脉病毒，番茄感染性绿化病毒和烟草嘎嘎声病毒），包括内部扩增对照。分析已发表引物的特性，以查看它们是否可用于多重RT-PCR。然后通过在cDNA合成过程中添加牛血清白蛋白，增加PCR延伸时间，降低PCR延伸温度并优化每对引物的浓度来优化三种靶标病毒和植物内部对照的扩增。这种优化产生了多重检测，其检测灵敏度与单纯RT-PCR相同。这种方法提供了一种简单而又方便的方法，可以直接使用已发布的引物而不是设计新的引物来进行多重分析。多重RT-PCR是检测马铃薯中多种病毒的灵敏且经济高效的方法，可用于检疫和认证计划。

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