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

    Fingerprint

    Transcription
    Reverse Transcription
    Amplification
    Messenger RNA
    Assays
    Real-Time Polymerase Chain Reaction
    Wilms' Tumor Genes
    Monitoring
    Tumors
    Genes

    Keywords

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

    ASJC Scopus subject areas

    • Clinical Biochemistry

    Cite this

    Real-time reverse transcription loop-mediated isothermal amplification for rapid and simple quantification of WT1 mRNA. / Morishita, Soji; Tani, Hidenori; Kurata, Shinya; Nakamura, Kazunori; Tsuneda, Satoshi; Sekiguchi, Yuji; Noda, Naohiro.

    In: Clinical Biochemistry, Vol. 42, No. 6, 04.2009, p. 515-520.

    Research output: Contribution to journalArticle

    Morishita, Soji ; Tani, Hidenori ; Kurata, Shinya ; Nakamura, Kazunori ; Tsuneda, Satoshi ; Sekiguchi, Yuji ; Noda, Naohiro. / Real-time reverse transcription loop-mediated isothermal amplification for rapid and simple quantification of WT1 mRNA. In: Clinical Biochemistry. 2009 ; Vol. 42, No. 6. pp. 515-520.
    @article{8fa86324139a44109281748718302a9b,
    title = "Real-time reverse transcription loop-mediated isothermal amplification for rapid and simple quantification of WT1 mRNA",
    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.",
    keywords = "Gene marker, Leukemia, Minimal residual disease (MRD) monitoring, Reverse transcription loop-mediated isothermal amplification (RT-LAMP), RT-PCR, Wilms' tumor gene (WT1)",
    author = "Soji Morishita and Hidenori Tani and Shinya Kurata and Kazunori Nakamura and Satoshi Tsuneda and Yuji Sekiguchi and Naohiro Noda",
    year = "2009",
    month = "4",
    doi = "10.1016/j.clinbiochem.2009.01.013",
    language = "English",
    volume = "42",
    pages = "515--520",
    journal = "Clinical Biochemistry",
    issn = "0009-9120",
    publisher = "Elsevier Inc.",
    number = "6",

    }

    TY - JOUR

    T1 - Real-time reverse transcription loop-mediated isothermal amplification for rapid and simple quantification of WT1 mRNA

    AU - Morishita, Soji

    AU - Tani, Hidenori

    AU - Kurata, Shinya

    AU - Nakamura, Kazunori

    AU - Tsuneda, Satoshi

    AU - Sekiguchi, Yuji

    AU - Noda, Naohiro

    PY - 2009/4

    Y1 - 2009/4

    N2 - 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.

    AB - 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.

    KW - Gene marker

    KW - Leukemia

    KW - Minimal residual disease (MRD) monitoring

    KW - Reverse transcription loop-mediated isothermal amplification (RT-LAMP)

    KW - RT-PCR

    KW - Wilms' tumor gene (WT1)

    UR - http://www.scopus.com/inward/record.url?scp=61849094245&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=61849094245&partnerID=8YFLogxK

    U2 - 10.1016/j.clinbiochem.2009.01.013

    DO - 10.1016/j.clinbiochem.2009.01.013

    M3 - Article

    VL - 42

    SP - 515

    EP - 520

    JO - Clinical Biochemistry

    JF - Clinical Biochemistry

    SN - 0009-9120

    IS - 6

    ER -