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实时定量逆转录聚合酶链反应检测K562/A02细胞系中bcr-abl融合基因mRNA水平

[Detection of bcr-abl fusion gene mRNA level in K562/A02 cell line by real-time quantitative RT-PCR].

作者信息

Chen Bao-An, Zhou Gui-Na, Cheng Jian, Qiao Chun, Wu Yu-Jie, Li Jian-Yong, Ding Jia-Hua, Gao Chong, Zhao Gang, Wang Jun, Bao Wen, Song Hui-Hui

机构信息

Department of Hematology, Southeast University Medical College, Nanjing 210009, Jiangsu Province, China.

出版信息

Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2011 Feb;19(1):40-3.

Abstract

This study was aimed to quantitatively analyze the mRNA level of bcr-abl fusion gene in K562/A02 cell line by real-time quantitative reverse transcriptase polymerase chain reaction (RQ-RT-PCR) technique. After being cultured for a period of time, the K562/A02 cell line was collected and RNA was extracted using TRIzoL kit. The real-time quantitative reverse transcriptase polymerase chain reaction technology was used to detect the level of bcr-abl fusion gene and internal reference abl gene. The results showed that a fine reproducibility was obtained between 10(7) and 10(3) copies/ml, reproducible sensitivity of RQ-RT-PCR was 10(-5). The expression of bcr-abl fusion gene in K562/A02 cells was higher and the level of bcr-abl mRNA was more than 100% in K562/A02 cells. It is concluded that RQ-RT-PCR is a reliable, sensitive and reproducible method for detecting mRNA level of bcr-abl fusion gene, which may be useful in monitoring the chronic myeloid leukemia.

摘要

本研究旨在通过实时定量逆转录聚合酶链反应(RQ-RT-PCR)技术定量分析K562/A02细胞系中bcr-abl融合基因的mRNA水平。培养一段时间后,收集K562/A02细胞系,使用TRIzol试剂盒提取RNA。采用实时定量逆转录聚合酶链反应技术检测bcr-abl融合基因和内参abl基因的水平。结果显示,在10(7)至10(3)拷贝/毫升之间获得了良好的重复性,RQ-RT-PCR的可重复灵敏度为10(-5)。K562/A02细胞中bcr-abl融合基因的表达较高,K562/A02细胞中bcr-abl mRNA水平超过100%。结论是,RQ-RT-PCR是检测bcr-abl融合基因mRNA水平的可靠、灵敏且可重复的方法,这可能有助于监测慢性粒细胞白血病。

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