Duplex reverse transcription-polymerase chain reaction system to detect Potato spindle tuber viroid using an internal control mRNA and a non-infectious positive control RNA
Duplex reverse transcription-polymerase chain reaction system to detect Potato spindle tuber viroid using an internal control mRNA and a non-infectious positive control RNA
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DOI：
10.1007/s10327-009-0159-z
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发表时间：
2009-06-01
影响因子：
1.2
通讯作者：
Hataya, Tatsuji
中科院分区：
文献类型：
--
作者：
Hataya, Tatsuji
基于双工逆转录聚合酶链式反应（RT-PCR）并结合植物 mRNA 内参，开发了一种准确可靠的马铃薯纺锤块茎类病毒（PSTVd）检测系统，以评估从休眠马铃薯块茎中提取的 RNA 的质量和 RT-PCR 的有效性。此外，在体外生成了基于 PSTVd 序列的非感染性 RNA，作为易于获得的阳性对照，且没有逃逸的可能性；该 RNA 与 PSTVd 感染中的真阳性有区别。该检测系统将在植物检疫和植物检疫认证应用中发挥作用。
An accurate and reliable system to detect Potato spindle tuber viroid (PSTVd) was developed based on duplex reverse transcription-polymerase chain reaction (RT-PCR) in combination with a plant mRNA internal control to evaluate the quality of RNA extracted from a dormant potato tuber and the effectiveness of the RT-PCR. In addition, a non-infectious RNA based on the PSTVd sequence was generated in vitro as a readily available positive control without the possibility of escape; this RNA was distinguishable from true positives in PSTVd infection. This detection system will be useful in plant quarantine and phytosanitary certification applications.