title: Integrated three-dimensional system-on-chip for direct quantitative detection of mitochondrial DNA mutation in affected cells.
abstract: We report a microfluidic system for automatic mitochondrial mutation diagnostics from sample purification to quantitative analysis. The system achieved direct DNA (mtDNA) mutation quantification in affected cells using a new 3D-microfluidic system, which integrated a mtDNA extraction module and a mutation detection module. Effective direct mtDNA extraction from the cells was realized using magnetic field manipulation. The obtained mtDNAs were subject to a fully automatic processing for quantitative mutation detection using integrated micropumps, micromixer and microtemperature control modules capable of mutation sensing by restriction enzyme digestion and real-time on-chip micro-PCR. Compared with traditional methods, this microfluidic system demonstrates the advantages of faster detection, requirement of fewer amount of specimens and reagents, much compact design and lower cost as well as lower risks for human errors. Thus, such system-on-chip would encourage the future translational development of rapid pathogenic mtDNA defects detection to provide more efficient clinical diagnosis and disease management strategies.
authors: Department of Engineering Science, National Cheng Kung University, Tainan, Taiwan.
authors: Chen-Min
authors: CM
authors: Chang
authors: Li-Fang
authors: LF
authors: Chiu
authors: Yau-Huei
authors: YH
authors: Wei
authors: Dar-Bin
authors: DB
authors: Shieh
authors: Gwo-Bin
authors: GB
authors: Lee
mesh_terms: D004252:DNA Mutational Analysis / Q000295:instrumentation*; D004272:DNA, Mitochondrial / Q000235:genetics* / Q000302:isolation & purification; D004867:Equipment Design; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D017354:Point Mutation; D060888:Real-Time Polymerase Chain Reaction / Q000295:instrumentation
chemical_list: D004272:DNA, Mitochondrial