Detection of bcr-abl fusion gene mRNA level in K562/A02 cell line by real-time quantitative RT-PCR.
- Author:
Bao-An CHEN
1
;
Gui-Na ZHOU
;
Jian CHENG
;
Chun QIAO
;
Yu-Jie WU
;
Jian-Yong LI
;
Jia-Hua DING
;
Chong GAO
;
Gang ZHAO
;
Jun WANG
;
Wen BAO
;
Hui-Hui SONG
Author Information
1. Department of Hematology, Southeast University Medical College, Nanjing 210009, Jiangsu Province, China. cba8888@hotmail.com
- Publication Type:Journal Article
- MeSH:
Fusion Proteins, bcr-abl;
genetics;
Humans;
K562 Cells;
RNA, Messenger;
genetics;
Real-Time Polymerase Chain Reaction;
methods;
Reverse Transcriptase Polymerase Chain Reaction;
methods;
Sensitivity and Specificity
- From:
Journal of Experimental Hematology
2011;19(1):40-43
- CountryChina
- Language:Chinese
-
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.
