Yuki Hashimoto1, Yuki Hatayama1, Nao Kojima1, Shota Morishita1, Satoko Matsumoto1, Yuzuru Hosoda2, Ayako Hara1, Toru Motokura3. 1. Division of Clinical Laboratory, Tottori University Hospital, Yonago 683-8504, Japan. 2. †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. 3. 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.
Abstract
BACKGROUND: Acute promyelocytic leukemia (APL) is a disease characterized by expression of Promyelocytic Leukemia-Retinoic Acid Receptor α (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 of PML-RARα mRNA in a single reaction. Serial dilutions of plasmid DNA containing bcr1, bcr2, or bcr3 PML-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. CONCLUSION: We developed an RT-LAMP assay for simple and rapid PML-RARα mRNA detection that may be clinically useful for point-of-care testing and APL diagnosis.
BACKGROUND:Acute promyelocytic leukemia (APL) is a disease characterized by expression of Promyelocytic Leukemia-Retinoic Acid Receptor α (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 of PML-RARα mRNA in a single reaction. Serial dilutions of plasmid DNA containing bcr1, bcr2, or bcr3 PML-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. CONCLUSION: We developed an RT-LAMP assay for simple and rapid PML-RARα mRNA detection that may be clinically useful for point-of-care testing and APL diagnosis.
Authors: J J van Dongen; E A Macintyre; J A Gabert; E Delabesse; V Rossi; G Saglio; E Gottardi; A Rambaldi; G Dotti; F Griesinger; A Parreira; P Gameiro; M G Diáz; M Malec; A W Langerak; J F San Miguel; A Biondi Journal: Leukemia Date: 1999-12 Impact factor: 11.528
Authors: Jerald P Radich; Edward Briercheck; Daniel T Chiu; Manoj P Menon; Olga Sala Torra; Cecilia C S Yeung; Edus H Warren Journal: Annu Rev Pathol Date: 2022-01-24 Impact factor: 32.350