Development of Reverse Transcription Loop-Mediated Isothermal Amplification for Simple and Rapid Detection of Promyelocytic Leukemia-Retinoic Acid Receptor ? mRNA.

ABSTRACT: Acute promyelocytic leukemia (APL) is a disease characterized by expression of Promyelocytic Leukemia-Retinoic Acid Receptor ? (PML-RAR?) 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).An RT-LAMP primer set was designed to detect three types of PML-RAR? mRNA in a single reaction. Serial dilutions of plasmid DNA containing bcr1, bcr2, or bcr3 PML-RAR? 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.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.We developed an RT-LAMP assay for simple and rapid PML-RAR? mRNA detection that may be clinically useful for point-of-care testing and APL diagnosis.

SUBMITTER: Hashimoto Y 

PROVIDER: S-EPMC5214692 | biostudies-literature | 2016 Dec

REPOSITORIES: biostudies-literature