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 × 109 copies and the threshold time with a correlation coefficient of R2 > 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.
- Gene marker
- Minimal residual disease (MRD) monitoring
- Reverse transcription loop-mediated isothermal amplification (RT-LAMP)
- Wilms' tumor gene (WT1)
ASJC Scopus subject areas
- Clinical Biochemistry