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

doi:10.1016/j.clinbiochem.2009.01.013

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^*

^*Corresponding author for this work

School of Advanced Science and Engineering

Research output: Contribution to journal › Article › peer-review

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 language	English
Pages (from-to)	515-520
Number of pages	6
Journal	Clinical Biochemistry
Volume	42
Issue number	6
DOIs	https://doi.org/10.1016/j.clinbiochem.2009.01.013
Publication status	Published - 2009 Apr

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

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10.1016/j.clinbiochem.2009.01.013

Cite this

@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, RT-PCR, Reverse transcription loop-mediated isothermal amplification (RT-LAMP), 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",

note = "Funding Information: This study was partially supported by the Global COE program “Center for Practical Chemical Wisdom” of the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan and financially supported by the Industrial Technology Research Grant Program in 2006 of the New Energy and Industrial Technology Development Organization (NEDO) of Japan.",

year = "2009",

month = apr,

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",

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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

N1 - Funding Information: This study was partially supported by the Global COE program “Center for Practical Chemical Wisdom” of the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan and financially supported by the Industrial Technology Research Grant Program in 2006 of the New Energy and Industrial Technology Development Organization (NEDO) of Japan.

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 - RT-PCR

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

KW - Wilms' tumor gene (WT1)

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

Abstract

Keywords

ASJC Scopus subject areas

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