title: [Detection of bcr-abl fusion gene mRNA level in K562/A02 cell line by real-time quantitative RT-PCR].
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.
authors.affiliation: Department of Hematology, Southeast University Medical College, Nanjing 210009, Jiangsu Province, China. cba8888@hotmail.com
authors.forename: Bao-An
authors.initials: BA
authors.lastname: Chen
authors.forename: Gui-Na
authors.initials: GN
authors.lastname: Zhou
authors.forename: Jian
authors.initials: J
authors.lastname: Cheng
authors.forename: Chun
authors.initials: C
authors.lastname: Qiao
authors.forename: Yu-Jie
authors.initials: YJ
authors.lastname: Wu
authors.forename: Jian-Yong
authors.initials: JY
authors.lastname: Li
authors.forename: Jia-Hua
authors.initials: JH
authors.lastname: Ding
authors.forename: Chong
authors.initials: C
authors.lastname: Gao
authors.forename: Gang
authors.initials: G
authors.lastname: Zhao
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
authors.forename: Wen
authors.initials: W
authors.lastname: Bao
authors.forename: Hui-Hui
authors.initials: HH
authors.lastname: Song
mesh_terms: D016044:Fusion Proteins, bcr-abl / Q000235:genetics*; D006801:Humans; D020014:K562 Cells; D012333:RNA, Messenger / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D012333:RNA, Messenger; D016044:Fusion Proteins, bcr-abl