title: Detection of BCR-ABL using one step reverse transcriptase- polymerase chain reaction and microchip electrophoresis.
abstract: One-step reverse transcriptase polymerase chain reaction (RT-PCR) coupled with microchip electrophoresis (MCE) was established to analyze BCR-ABL fusion gene. The use of one-step RT-PCR could simplify the RT-PCR procedure and thus reduced the risk of contamination and sample consumption. This method also enhanced the sensitivity for amplified target DNA and dramatically shorted the analysis time. Moreover, this assay can simultaneously identify b2a2 and b3a2. Orthogonal array design, which can investigated mutual effects of PCR parameters, was used to optimize the reaction system. This approach was highly effective, reproducible and sensitive, and would be suitable for the determination of BCR-ABL in clinic diagnosis.
authors.affiliation: State Key Laboratory of Chemical Resource Engineering, Beijing University of Chemical Technology, Beijing 100029, China; Department of Chemistry, Beijing Key Laboratory of Microanalytical Methods and Instrumentation, Tsinghua University, Beijing 100084, China.
authors.forename: Xuexia
authors.initials: X
authors.lastname: Lin
authors.forename: Jing
authors.initials: J
authors.lastname: Wu
authors.forename: Wu
authors.initials: W
authors.lastname: Liu
authors.forename: Haifang
authors.initials: H
authors.lastname: Li
authors.forename: Zhihua
authors.initials: Z
authors.lastname: Wang
authors.forename: Jin-Ming
authors.initials: JM
authors.lastname: Lin
mesh_terms: D046209:Electrophoresis, Microchip; D016044:Fusion Proteins, bcr-abl / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D020014:K562 Cells; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000175:diagnosis / Q000235:genetics; D025202:Molecular Diagnostic Techniques; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity
chemical_list: D016044:Fusion Proteins, bcr-abl
keywords: Microchip electrophoresis; One-step RT-PCR; Orthogonal array design