title: Development of Reverse Transcription Loop-Mediated Isothermal Amplification for Simple and Rapid Detection ofPromyelocytic Leukemia-Retinoic Acid Receptorα mRNA.
abstract: BACKGROUND
  Acute promyelocytic leukemia (APL) is a disease characterized by expression ofPromyelocytic Leukemia-Retinoic Acid Receptorα (PML-RARα) chimeric mRNA. Although APL is curable, early death due to hemorrhage is a major problem. Here, we report the development of a simple and rapid diagnostic method for APL based on reverse transcription loop-mediated isothermal amplification (RT-LAMP).
  
  
  METHODS
  An RT-LAMP primer set was designed to detect three types ofPML-RARα mRNA in a single reaction. Serial dilutions of plasmid DNA containing bcr1, bcr2, or bcr3PML-RARα sequences and RNA extracted from bone marrow aspirates of 6 patients with APL were used to compare the results of RT-LAMP and nested PCR assays.
  
  
  RESULTS
  Plasmid DNA was amplified by RT-LAMP, for which the reaction time was > 4 h shorter and the lower detection limit was higher than for nested RT-PCR. Six of 7 samples tested positive by both methods.
  
  
  CONCLUSIONS
  We developed an RT-LAMP assay for simple and rapidPML-RARα mRNA detection that may be clinically useful for point-of-care testing and APL diagnosis.
authors.affiliation: Division of Clinical Laboratory, Tottori University Hospital, Yonago 683-8504, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Hashimoto
authors.affiliation: Division of Clinical Laboratory, Tottori University Hospital, Yonago 683-8504, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Hatayama
authors.affiliation: Division of Clinical Laboratory, Tottori University Hospital, Yonago 683-8504, Japan.
authors.forename: Nao
authors.initials: N
authors.lastname: Kojima
authors.affiliation: Division of Clinical Laboratory, Tottori University Hospital, Yonago 683-8504, Japan.
authors.forename: Shota
authors.initials: S
authors.lastname: Morishita
authors.affiliation: Division of Clinical Laboratory, Tottori University Hospital, Yonago 683-8504, Japan.
authors.forename: Satoko
authors.initials: S
authors.lastname: Matsumoto
authors.affiliation: †Department of Hematology, Tottori University Hospital, Yonago 683-8504, Japan; ‡Division of Clinical Laboratory Medicine, Department of Pathophysiological and Therapeutic Science, School of Medicine, Tottori University Faculty of Medicine, Yonago 683-8503, Japan.
authors.forename: Yuzuru
authors.initials: Y
authors.lastname: Hosoda
authors.affiliation: Division of Clinical Laboratory, Tottori University Hospital, Yonago 683-8504, Japan.
authors.forename: Ayako
authors.initials: A
authors.lastname: Hara
authors.affiliation: Division of Clinical Laboratory, Tottori University Hospital, Yonago 683-8504, Japan; †Department of Hematology, Tottori University Hospital, Yonago 683-8504, Japan; ‡Division of Clinical Laboratory Medicine, Department of Pathophysiological and Therapeutic Science, School of Medicine, Tottori University Faculty of Medicine, Yonago 683-8503, Japan.
authors.forename: Toru
authors.initials: T
authors.lastname: Motokura
keywords: Promyelocytic Leukemia–Retinoic Acid Receptor α; acute promyelocytic leukemia; polymerase chain reaction; reverse transcription loop-mediated isothermal amplification; t(15;17) chromosome translocation