[Quantification of PML/RAR alpha fusion gene transcripts in patients with acute promyelocytic leukemia by using real-time quantitative PCR].
OBJECTIVE
To establish and evaluate a real time quantitative PCR (RQ-PCR) method for detection and quantification the PML/RAR alpha fusion gene transcripts in patients with acute promyelocytic leukemia (APL).


METHODS
Three pairs of primers and TaqMan probe were designed for detecting the most frequent PML/RAR alpha transcripts (L-form, S-form and V-form) and normal ABL was used as an internal control. A real time PCR condition was established to detect PML/RAR alpha and ABL positive templates with a series of dilutions. To evaluate this assay, bone marrow samples from 6 APL patients were detected.


RESULTS
In repeated tests, maximal sensitivities of 10 copies/microL were obtained, while reproducible maximal sensitivity achieved 100 copies/microL. In 10 normal controls, no amplified fluorescent signals were detected. The median absolute and normalized amount of PML/RAR alpha fusion gene transcripts were 4.27 x 10(3)-3.36 x 10(5) copies/50 ng (median 4.33 x 10(4)copies/50 ng) and 29.38%-600.53% (median 48.12%) respectively. One case showed significant decrease of PML/RAR alpha fusion gene transcripts after induction therapy compared to that at the time of diagnosis, while the fusion transcripts significantly increased after relapsed.


CONCLUSIONS
RQ-PCR is a sensitive, reliable quantitative assay and can be used in the diagnosis of APL and measurement of MRD.
Key Laboratory of Experimental Hematolgy, Affiliated Peopleos Hospital, Jiangsu University, Zhenjiang, Jiangsu, 212002 People's Republic of China.
Jiang
J
Lin
Lan-xiu
LX
Han
Jun
J
Qian
Ya-li
YL
Wang
Zhen
Z
Qian
Xiao-fei
XF
Yang
Xiao-jing
XJ
Sheng
D000293:Adolescent; D000328:Adult; D005260:Female; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute / Q000175:diagnosis / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D015514:Oncogene Proteins, Fusion / Q000235:genetics*; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D055815:Young Adult
D015514:Oncogene Proteins, Fusion; C093476:promyelocytic leukemia-retinoic acid receptor alpha fusion oncoprotein