Real-time reverse transcription loop-mediated isothermal amplification for rapid and simple quantification of WT1 mRNA

Soji Morishita, Hidenori Tani, Shinya Kurata, Kazunori Nakamura, Satoshi Tsuneda, Yuji Sekiguchi, Naohiro Noda

Research output: Contribution to journalArticle

10 Citations (Scopus)

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 × 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.

Original languageEnglish
Pages (from-to)515-520
Number of pages6
JournalClinical Biochemistry
Volume42
Issue number6
DOIs
Publication statusPublished - 2009 Apr 1

    Fingerprint

Keywords

  • Gene marker
  • Leukemia
  • Minimal residual disease (MRD) monitoring
  • RT-PCR
  • Reverse transcription loop-mediated isothermal amplification (RT-LAMP)
  • Wilms' tumor gene (WT1)

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this