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.
Methods. 2013 Dec 15;64(3):250-4. doi: 10.1016/j.ymeth.2013.05.010. Epub 2013 Jun 6.
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.
建立了一步法逆转录聚合酶链反应（RT-PCR）结合微芯片电泳（MCE）分析 BCR-ABL 融合基因。一步法 RT-PCR 的使用可以简化 RT-PCR 程序，从而降低污染和样品消耗的风险。该方法还提高了扩增靶 DNA 的灵敏度，并大大缩短了分析时间。此外，该测定法还可以同时鉴定 b2a2 和 b3a2。正交数组设计可用于研究 PCR 参数的相互影响，以优化反应体系。这种方法非常有效、可重复且灵敏，适用于临床诊断中 BCR-ABL 的测定。
