OBJECTIVES This study developed a novel MRD monitoring method targeting Wilms' tumor gene (WT1) mRNA using reverse transcription loop-mediated isothermal amplification (RT-LAMP). DESIGN AND METHODS A primer set for the assay was designed on the basis of the sequences between the 17AA and KTS regions of WT1mRNA. WT1 mRNA was quantified by real-time RT-LAMP and the accuracy of RT-LAMP was compared with that of real-time RT-PCR. RESULTS The standard curve was expressed as a linear relationship between the log copy numbers of WT1 mRNA ranging from 6.8 x 10 to 6.8 x 10(9) copies and the threshold time with a correlation coefficient of R(2) > 0.994. The measured values obtained by RT-LAMP strongly correlated with those obtained by real-time RT-PCR. CONCLUSION RT-LAMP can be used to determine WT1 mRNA expression levels. This assay will contribute to a more specific, simple, and rapid MRD monitoring than conventional assays.

中文翻译：

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实时逆转录环介导的等温扩增，用于快速，简单地定量WT1 mRNA。

目的本研究开发了一种新的MRD监测方法，该方法使用逆转录环介导的等温扩增（RT-LAMP）靶向Wilms的肿瘤基因（WT1）mRNA。设计与方法根据WT1mRNA的17AA和KTS区之间的序列设计了用于该测定的引物组。通过实时RT-LAMP对WT1 mRNA进行定量，并将RT-LAMP的准确性与实时RT-PCR的准确性进行比较。结果标准曲线以WT1 mRNA的对数拷贝数从6.8 x 10到6.8 x 10（9）拷贝与阈值时间之间的线性关系表示，相关系数R（2）> 0.994。通过RT-LAMP获得的测量值与通过实时RT-PCR获得的测量值高度相关。结论RT-LAMP可用于确定WT1 mRNA表达水平。