Abstract
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 WT1 mRNA. 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 × 10 to 6.8 × 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.
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