Real-time quantitative RT PCR detection of acute promyelocytic leukemia PML RARa fusion gene.docVIP

 PAGE \* MERGEFORMAT 17 Real-time quantitative RT PCR detection of acute promyelocytic leukemia PML RARa fusion gene Author: DUAN Lian-Ning Lang Li Li Huimin Liu Yi-Min Wang Xue-Mei Zhang Abstract This study was designed to establish real-time quantitative RT  PCR detection of acute promyelocytic leukemia (APL) PML  RARa mRNA way to explore the APL PML  RARa fusion gene expression and effect relationship. RT  PCR amplification using cultured NB4 cells, PML  RARa fusion gene, taking 1μg NB4 cells for 10 times the total cDNA gradient dilution, as the standard products, the establishment of fluorescence quantitative RT  PCR method, the method sensitivity, stability , repeatability were determined. Quantitative detection of four cases of acute promyelocytic leukemia (APL) patients before and after treatment of bone marrow PML  RARa fusion gene transcript levels, and one cases of relapse after complete remission after treatment in patients with PML  RARa transcript level (copy number) dynamic monitoring. The results showed that: The present study to establish a real-time quantitative RT  PCR method can detect 10-5 μg NB4 cell cDNA of PML  RARa fusion gene, its repetitive CT value, control rooms, and inter-coefficient of variation (CV), respectively 2.1%, 3.8%. 4 cases of bone marrow in patients with newly diagnosed PML  RARa gene transcript levels were 1 884,5 533,1 803,4 677 copies, with an average of 3475 copies. After treatment by ATRA chemotherapy PML  RARa gene transcription levels fall to 40,135,79,229 copy, with an average of 121 copies. There are one cases of patients before treatment PML  RARa gene transcript level of 8600 copies, after 4 months of treatment, although the patient remains in complete remission at this time, but its level of transcription copies still 730.3 months after the patients recurrence, transcript levels rose to 11 000 copies. After ATRA with chemotherapy in this post-transcriptional level also gradually decreas